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.     

A Model of the Relationship Between Psychosocial Variables and Diurnal Cortisol Rhythm Under Chronic Stress by Using Structural Equations

By using structural equations, we investigated the effect of chronic stress on salivary cortisol rhythm and proposed a causal model of chronic stress by using psychosocial and physiological data. First, 111 healthy workers (48 males, 63 females) completed questionnaires on chronic stress and lifestyle habits. Then, they provided saliva samples and answered questionnaires that were prepared to assess their psychological states 5 times (on waking up and at 10:00, 11:40, 14:00, and 16:00) on workdays. Structural equation modeling (SEM) revealed that chronic stress and longer commuting time resulted in sleep irregularities and this disrupted the cortisol circadian rhythm. This suggests that chronic stress disrupts the cortisol circadian rhythm even in healthy individuals, and sleep regularity mediates the effect of chronic stress on the cortisol rhythm.

     

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.     

Air vent in water chamber and surface coating on liner slides concerning auxiliary cooling system for the high temperature engineering test reactor

Experimental study was made to confirm the validity of new designs of the auxiliary cooling system for the high temperature engineering test reactor (HTTR). First, it is necessary to vent residence air in outlet side of water chamber of the auxiliary heat exchanger for the HTTR. Accordingly, we have proposed to mount a proper bend duct in the outlet side of the water chamber. Air vent is done by difference between pressures at both ends of the bend duct caused by the forced water circulation using the water pumps. From flow tests, it was confirmed that it is capable of venting the air through the bend duct by circulating the water in maximum capacity of the water pumps. Second, it is essential to prevent seizure and excessive wear of the liner slides of the auxiliary concentric hot gas duct for the HTTR at a service temperature of 950°C. Therefore, we have put forward to coat titanium nitride (TiN) on the surface of the liner slides made of nickel-based superalloy Hastelloy XR using the thermochemical vapor deposition method. As a result of seizure and wear tests, it was confirmed that the TiN coating film of 3 μm on the surface of Hastelloy XR is sufficient.     

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     

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.     

Determination of assimilable organic carbon (aoc) in ozonated water with acinetobacter calcoaceticus

The effect of ozone application in drinking water on the production of assimilable organic carbon (AOC) was evaluated. The typical procedure to determine AOC is suggested by van der Kooij, which is the method of bacterial growth measurement by colony‐forming units using the strain P17 and/or NOX. The bacterial indicator species used for this study is Acinetobacter calcoaceticus which was isolated and identified while ozonating Nakdong river water. This strain could never be isolated from the raw water, but this strain was the predominant isolate in the ozonated water. Within a short incubation time, this organism was found to replicate well on acetate and oxalate as the sole carbon sources. The yield coefficients of this organism for acetate and oxalate are the same order of magnitude as the value of P17 and NOX. With full‐scale experiments, A. calcoaceticus concentration was found to increase after ozonation, but did not decrease upon chlorination. In laboratory‐scale experiments with Yongsan river water, aldehyes were found to be produced in proportion to the ozone dose. The raw water contains low concentrations of aldehydes, but has a high AOC concentration. A correlation between aldehyde production and AOC production was observed in the tested water with ozonation.