Pinpoint injection of microtools for minimally invasive micromanipulation of microbe by laser trap

This paper reports transportation of the target microbe by the laser trapped microtools with minimum laser irradiation to the target. The size of a microtool (MT) is around micrometer. The MTs are manipulated by the focused laser under the microscope to manipulate the target microbe. Here we propose a pinpoint injection method of MTs at the desired location in the microchamber, which is filled with liquid. At first, we classified the injection method of the MTs in four categories. Here we employed a new method to install the MTs inside the microchamber. We developed a MT holding chip to install the MTs. The MTs were injected in the microchamber, and were manipulated successfully by the laser scanning micromanipulator to transport the target microbe for separation. The proposed method is useful for the pinpoint injection of MTs and separation by the indirect micromanipulation.     

Construction of a plasmid carrying both CTP synthetase and a fused gene formed from cholinephosphate cytidylyltransferase and choline kinase genes and its application to industrial CDP-choline production: enzymatic production of CDP-choline from orotic acid (Part II)

A new method for enzymatic production of cytidine diphosphate choline (CDP-choline) from orotic acid and choline chloride was developed. To establish an industrial manufacturing process, we constructed a plasmid, pCKG55, which simultaneously expressed in Escherichia coli the three following enzymes; CTP synthetase (encoded by the pyrG gene from E. coli), cholinephosphate cytidylyltransferase (encoded by the CCT gene from Saccharomyces cerevisiae), and choline kinase (encoded by the CKI gene from S. cerevisiae). CCT and CKI genes on pCKG55 were designed to be expressed as a single CCT/CKI fused protein. This CCT/CKI fused protein retained both activities and the thermal stability of its cholinephosphate cytidylyltransferase activity was nearly the same as the native CCT enzyme. Corynebacterium ammoniagenes KY13505 and E. coli MM294/pCKG55 were cultured in 5-liter jar fermentor independently. Equal volumes of each broth were mixed in a 2-liter jar fermentor, and then the enzymatic reaction was done using 47 mM orotic acid and 60 mM choline chloride as substrates. After 23 h of the reaction at 32 degrees C, 21.5 mM (11 g/liter) of CDP-choline was accumulated.     

Construction of the plasmid PMEX8-HAK1 and random site-directed mutagenesis of human cytosolic adenylate kinase

The pMEX8-hAK1 vector was devised from the pAK plasmid (Kim J. H. et al., 1989, Protein Engineering 5, 379-386), which could directly express human adenylate kinase proteins without recombination and its single strand DNA could be withdrawn with helper phage for random site-directed mutagenesis. The conserved key residues at Lys21, Lys27, and Thr39 were engineered to obtain mutants for kinetic analysis. Three mutants were obtained as K21P, K27R, and T39S, their specific activities were strikingly reduced compared to those of wild type adenylate kinase. This pMEX8-hAK1 will be a powerful tool for site-directed mutagenesis to detect the substrate-enzyme interaction for human adenylate kinase including various other enzymes.     

Impulse breakdown voltages of triggered multistage vacuum gap

Several sealed-off triggered vacuum gaps are connected in series to improve hold-off voltage. The characteristics of impulse breakdown voltage of these series-connected gaps are investigated experimentally. The sum hold-off voltage of series-connected gaps decreases to a unit hold-off voltage when the maximum value of voltage division ratio across the gaps increases to unity. Self-breakdown probability of the series-connected gaps is always higher than that of a single gap under the same conditions. Hence, stage efficiency of the multistage gap decreases with increasing number of stages. Its value is 90 percent with 2-stage gap and 75 percent with 5-stage gap, respectively, under the same voltage division ratio and the same gap length (2.0 mm) in each stage. Triggered breakdown voltage of 2- or 3- stage gap is several hundred volts when all gaps are triggered simultaneously at the peak of the main impulse wave and a working voltage range is nearly 100 percent in this case. The working voltage range decreases with number of stages. Its value is 45 percent with 3-stage gap and 15 percent with 5-stage gap, respectively, when one triggered gap is fired for switching.     

The temperature dependence of luminescence from a long-lasting phosphor exposed to ionizing radiation

The temperature dependence of luminescence from a long-lasting phosphor (LLP), SrAl₂O₄ : Eu²⁺,Dy³⁺, exposed to ionizing radiation has been measured to understand the LLP luminescence mechanism. Evaluation of the decay constants of the LLP exposed to -, β- or γ-rays at temperatures from 200 to 390 K showed that the decay constant is divided into four components ranging from 10⁻⁴ to 10⁻¹ s⁻¹ with activation energies of 0.02–0.35 eV.

Total luminous intensity from the LLP with changing irradiation temperature has its maximum value around the room temperature. Irradiation at elevated temperature (390 K) has the total luminescence pattern with monotonous decrease as temperature rises. As a result of evaluating the temperature dependence of luminescence, the luminescence mechanism is considered as follows:

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A high-speed halftoning processor for raster scanned images

A high-speed bilevel reproduction algorithm, called modified error diffusion (MED) algorithm, has been developed to provide high-quality halftoning images for continuous tone images and has been implemented in a CMOS LSI chip. The chip has been designed with standard-cell 1.5-μm CMOS technology using an optimum layout design. The chip has achieved a maximum processing speed of 60 ns/pixel     

Extensions of polyacrylic acid ammonium salts in the adsorption layer to fluidize alumina slurries

An average shell volume occupied in the adsorption layer on alumina by a polyacrylic acid ammonium salts molecule (PAA) defined as the average area occupied by an adsorbed PAA on alumina multiplied by the average thickness of the water layer at the limit of thickening, has been calculated from the adsorbed amount of PAA and from the flow points of alumina in the presence of PAA of different molecular weights. A steric effect of the PAA dispersant on the dispersion of alumina resulted in a flow with no yield stress. This was due to the change of the extension of PAA in the adsorption shell from a sphere to an ellipsoid with a long axis which exceeded the effective distance that the van der Waals attraction force reaches at a molecular weight for the PAA of between 10 000–20 000.     

Physical and functional association of cortactin with Syk in human leukemic cell line K562

Human leukemic cell line K562 is induced to differentiate into the megakaryocytic lineage by stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA). We demonstrate here that TPA stimulation increases tyrosine phosphorylation of an 80-kDa protein at an early stage of megakaryocytic differentiation and that this 80-kDa protein is identical with cortactin. Since tyrosine kinase Syk was activated by TPA stimulation, we examined the possibility that cortactin is a potential substrate of Syk in K562 cells. TPA-induced tyrosine phosphorylation of cortactin was decreased profoundly by overexpression of dominant-negative Syk. Furthermore, cortactin was associated with Syk even before TPA stimulation. Since cortactin was previously referred as an 80/85-kilodalton pp60src substrate, we examined the association between Src and cortactin, whereas its association could not be detected. These data suggest that Syk phosphorylates cortactin in K562 cells upon TPA treatment.     

High-power CW-DFB laser diode modules for CATV applications operating at 1550 nm

Investigation of the correlation between longitudinal photon density distribution and spectral linewidth re-broadening, in conjunction with carefully designed coupling optics, enable laser modules that simultaneously achieve very high fibre-coupled power of 175 mW and very narrow linewidth<1 MHz even at /spl sim/120 mW of output power to be successfully fabricated.