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 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which could directly express human adenylate kinase proteins without recombination and its single strand DNA could be withdrawn with helper phage for random site-directed mutagenesis.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.     

Enhancement by ganglioside GT1b of annexin I phosphorylation in bovine mammary gland in the presence of phosphatidylserine and Ca2+

Ganglioside GT1b and, to a lesser extent, GD3, enhanced phosphorylation of a 36 kDa protein (the substrate of protein kinase C) in the particulate fraction from bovine mammary gland. Sialic acids, asialogangliosides, and GM3 were without effect, and GD1a conversely inhibited phosphorylation of the 36 kDa protein. The enhanced phosphorylation by GT1b required the simultaneous presence of phosphatidylserine (PS) and Ca²⁺. The 36 kDa protein reacted with anti-annexin I in Western blot analysis. Addition of purified annexin I to the reaction mixture containing the particulate fraction increased the extent of phosphorylated 36 kDa protein, and the phosphorylation was further enhanced by GT1b. The enhanced phosphorylation of annexin I by GT1b was also dependent on PS and Ca²⁺. When annexin I was phosphorylated by purified protein kinase C, GT1b inhibited the annexin I phosphorylation. Addition of epidermal growth factor or insulin to the particulate fraction had little effect on the enhancement. These results suggest that an enzyme or enzymes other than protein kinase C, epidermal growth factor receptor kinase, or insulin receptor kinase is responsible for the GT1b- and GD3-enhanced phosphorylation of annexin I in the presence of PS and Ca²⁺.     

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     

Fundamental solutions for half plane with an oblique edge crack

An elastic half plane with an oblique edge crack is considered in this paper. A pair of concentrated forces or point dislocations is assumed to act at an arbitrary point in the half plane. The half plane with an edge crack is first mapped into a unit circle by a rational mapping function so that the following analysis can be carried out on the mapped plane analytically. Then the complex stress functions are derived by separating the whole problem into two parts; one is the principal part corresponding to the infinite plane acted on by concentrated forces or dislocations, the other is the holomorphic part, which can be determined by making use of the property of regularity of complex stress functions. The stress intensity factors of the crack can be calculated with different inclined angles of the crack, and the displacement and stress components at an arbitrary position in the half plane can be expressed explicitly.     

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.     

The effects of R-75,317 on antiglomerular basement membrane glomerulonephritis in rats

Platelet-activating factor (PAF) is a potent inflammatory mediator which is released by various inflammatory cells and produced by certain tissues, including the kidney. PAF has been shown to increase glomerular permeability to protein and to decrease glomerular filtration rate (GFR) by contracting mesangium. On the basis of these observations, it has been suspected that PAF may play a role as mediator of glomerular damage in glomerular nephritis. To examine this possibility, we studied the effects of a specific PAF antagonist, R-75,317, on the development of an experimental model of anti-glomerular basement membrane (anti-GBM) glomerulonephritis. Glomerulonephritis was initiated by injecting rabbit anti-rat GBM serum into rats. Proteinuria gradually developed after serum injection, plateaued at week 2, and remained at the high level of week 2 throughout the experimental period (6 wk). Chronic treatment with R-75,317 (10 mg/kg/day i.p.) tended to delay the onset of proteinuria and significantly accelerated the recovery phase. Creatinine clearance (Ccr) fell to 40% at week 3. R-75,317 treatment completely prevented this decline of Ccr. Histological changes in this model (glomerular hypertrophy, proliferation of mesangial matrix and interstitial fibrosis) were also ameliorated by the R-75,317 treatment. The results suggest that PAF may play a role in the development of glomerulonephritis and that PAF antagonists could be used in the treatment of human renal disease. Based on a paper presented at the Third International Conference on Platelet-Activating Factor and Structurally Related Alkyl Ether Lipids, May 1989.     

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.     

An image processing system using Image Signal Multiprocessors (ISMPs)

This paper describes an image processing system using Image Signal Multiprocessors (ISMPs) adapted to gray-level image preprocessing for image analysis and image enhancement. It is composed of four ISMPs, five 1H-delay-lines, two 512×512×8-bit frame memories, a video timing controller (VTC), two 256-word ×8-bit ×8-table Look Up Tables (LUTs) and 80 nsec/sampling A/D and D/A converters. This multiprocessor system performs convolution operations such as spatial filters, contrast enhancement, and binarization for gray-level images, thinning, thickening, pattern matching etc. for binary images, and image quality improvement for moving images such as T.V. images. Otherwise, it performs feature extraction operations such as area calculations, fillet coordination, and moment calculations for objective image data. Moreover, this system is capable of applying color image processing by using a multiboard system.