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.     

Augmentation of boiling heat transfer from horizontal cylinder to liquid by movable particles

This paper presents a series of experimental results on a passive augmentation technique of boiling heat transfer by supplying solid particles in liquid. A cylindrical heater 0.88 mm in diameter is placed in saturated water, in which a lot of mobile particles exist, and the nucleate and film boiling heat transfer characteristics are measured. Particle materials used were alumina, glass, and porous alumina, and the diameter ranged from 0.3 mm to 2.5 mm. Particles are fluidized by the occurrence of boiling without any additive power, and the heat transfer is augmented. The maximum augmentation ratio obtained in this experiment reaches about ten times the heat transfer coefficient obtained in liquid alone. The augmentation ratio is mainly affected by the particle material, diameter, and the height of the particle bed set at no boiling condition. The augmentation mechanism is discussed on the basis of the experimental results. © 2001 Scripta Technica, Heat Trans Asian Res, 31(1): 28–41, 2002     

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.     

Graphite formation on Ni film by chemical vapor deposition

Thin film formation of graphite by chemical vapor deposition using 2-methyl-1,2′-naphthyl ketone as a starting material was carried out on Ni film substrates. On Ni films directly deposited on quartz glass, the graphite films were obtained when the Ni film thickness was above 1 000 Å and above 5 000 Å at 700 °C and 1 000 °C, respectively. Depositions on thinner Ni film substrates comprise amorphous carbon (a-C) or graphite tubes which was owing to the thermal coagulation of the Ni film into droplets. On the other hand, graphite film was obtained on the Ni film with thickness 10 Å when a-C was inserted between the Ni film and the quartz glass. The coagulation of the Ni film is considered to be avoided by inserting a-C layer.     

Permeation of Zidovudine and Probenecid from Oily Bases Containing Alcohols Through Rat Skin

Permeation of zidovudine (3'-azido-3'-deoxythymidine, AZT) and probenecid from oily bases containing an alcohol through rat skin was examined. Isopropyl myristate (IPM), as an oily vehicle, showed a penetration enhancing effect for AZT and probenecid. Ethanol, n-propanol, and n-butanol were used as additives in IPM and were examined for their own permeation and the enhancing effect on the permeation of AZT and probenecid. The skin permeation of AZT and probenecid from IPM was enhanced by addition of the alcohol in IPM. The degree of the enhancement was decreased with increasing lipophilicity of the alcohol used. me permeation rate of the drug from those systems was shown to be governed by penetration-enhancing effects of the oily base and alcohol, and the penetration of the alcohol itself through the skin.     

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.     

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