Direct evidence of high DNA binding activity of transcription factor AP-1 in rheumatoid arthritis synovium

Green fluorescent protein (GFP) is increasingly being used in plant biology from the cellular level to whole plant level. At the cellular level, GFP is being used as an in vivo reporter to assess frequency of transient and stable transformation. GFP has also proven to be an invaluable tool in monitoring trafficking and subcellular localization of protein. At the organ level and up, many exciting applications are rapidly emerging. The development of brighter GFP mutants with more robust folding properties has enabled better macroscopic visualization of GFP in whole leaves and plants. One interesting example has been the use of GFP to monitor virus movement in and among whole plants. GFP is also emerging as a powerful tool to monitor transgene movement and transgenic plants in the field. In a proof-of-concept study, tobacco was transformed with a modified version of the GFP gene controlled by a constitutive (35S) promoter. GFP expression in progeny plants ranged from 0% to 0.5%, and approximately 0.1% GFP was the minimal amount needed for unambiguous macroscopic detection. GFP is the first truly in vivo reporter system useful in whole plants, and we project its usefulness will increase even further as better forms of GFP genes become available.     

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which could directly express human adenylate kinase proteins without recombination and its single strand DNA could be withdrawn with helper phage for random site-directed mutagenesis.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

Intracoronary infusion of E6010 has more potent thrombolytic activity than tissue plasminogen activator (t-PA) in dogs: a higher plasma level of E6010 than t-PA causes potent thrombolytic activity

We examined the thrombolytic properties of a novel modified human tissue plasminogen activator (PA) (E6010), in which cysteine 84 is replaced by serine, and which has a prolonged biologic half-life (t1/2). We compared the thrombolytic efficacy of continuous intracoronary (i.c.) infusion of E6010 with that of recombinant human tissue PA (rt-PA) in a canine model with copper coil-induced 1-h-old coronary artery thrombi and also compared the relation between thrombolytic efficacy and plasma clearance represented by pharmacokinetic parameters of i.c.-infused E6010 and rt-PA. Sixty-minute E6010 and rt-PA i.c. infusions were compared. The thrombolytic effects of i.c.-infused E6010 and rt-PA, represented by time to reperfusion (TR), reperfusion rate at 60 min (RR), and reocclusion rates at 60 min after reperfusion (OR) were as follows. E6010: Dose 0.06, 0.15, 0.3 (mg/kg/h); TR 25 +/- 10, 15 +/- 10, 13 +/- 5 (min); RR 100, 100, 100 (%); and OR 0, 0, 17 (%), respectively. Recombinant t-PA: Dose 0.06, 0.15, 0.3 (mg/kg/h); TR 47 +/- 12, 18 +/- 17, 14 +/- 4 (min); RR 50, 75, 100 (%); and OR 100, 33, 33 (%), respectively. These findings indicate that E6010 has more potent thrombolytic activity than rt-PA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Athermal silica-based arrayed-waveguide grating multiplexer

A temperature dependent channel wavelength shift in a silica-based arrayed-waveguide grating multiplexer is successfully suppressed from 0.95 to 0.05 nm in the 0-85°C temperature range, which means that it can be used in practical WDM systems without the need for temperature control     

Bromine addition and successive amine substitution of mesoporous ethylenesilica: Reaction, characterizations and arsenate adsorption

Bromination and subsequent ethylenediamine substitution of the CC double bond in mesoporous ethylenesilica were carried out to explore the characteristics of this periodic mesoporous organosilica. The structures of the products (BrPMO and EDA–BrPMO, respectively) were analysed by IR, Br K-edge EXAFS and NMR spectroscopies, as well as X-ray diffraction and nitrogen adsorption. We showed (1) that the formulae of the two products that formed were [CHBrSiO_1.5]_0.45[CHSiO_1.5]_0.55 and [NH₂CH₂CH₂NHCHSiO_1.5]_0.05 [CHBrSiO_1.5]_0.40[CHSiO_1.5]_0.55, respectively, (2) that the addition of Br₂ at room temperature occurred on the CC double bonds with disturbing the framework structure, (3) that IR absorption band of CC bonds that reacted with Br₂ is significantly different from that of inactive CC bond, (4) that the length of the C–Br bond was considerably longer than in conventional alkyl bromides, and (5) that a large proportion of the ν(C–Br) band remained at the same position in the IR absorption spectrum after the ethylenediamine (EDA) substitution, while a new ν(C–Br) absorption also appeared. The mechanisms of these reactions are discussed at both the micro and mesoscopic levels.

Arsenate adsorption on EDA–BrPMO, in which the EDA is directly bound to the “surface” of the mesopores, was compared with adsorption on EDA–Pr–PMO, which was prepared by the direct synthesis of 3-chloropropyl-functionalized mesoporous ethanesilica followed by the substitution of Cl with EDA. The strength of the adsorption, as measured with the distribution coefficient, was greater for the former adsorbent than the latter. The origin of this difference was attributed to the distance between amino group and the surface.     

Effective renaturation of reduced lysozyme by gentle removal of urea

To increase the folding yield of concentrated reduced lysozyme,we developed a renaturation method by means of dialysis fromconcentrated urea with redox agents. After lysozyme was incubatedin the reducing buffer (8 M urea solution) with oxidized glutathione,renaturation of reduced lysozyme was started by dialysis againstthe dialyzing buffer containing 8 M urea with redox agents.The urea concentration of the dialyzing bottle was graduallydiluted with dialyzing buffer without urea at a flow rate of0.1 ml/min by high pressure pump. Using this systematic dialysis,a concentration as high as 5 mg/ml of reduced lysozyme couldbe renaturated in 80% yield, while the folding yield was <5%even at a concentration of 1 mg/ml using a conventional rapiddilution method [Goldberg et al. (1991) Biochemistry, 30, 2790–2797].Therefore, it was concluded that gentle removal of urea fromdenatured proteins, dissolved in concentrated urea solution,by means of dialysis should be useful to renature denaturedproteins effectively.     

Catalyst‐Free GaN Nanorods Synthesized by Selective Area Growth

GaN nanorod formation on Ga‐polar GaN by continuous mode metalorganic chemical vapor deposition selective area growth (MOCVD SAG) is achieved under a relatively Ga‐rich condition. The Ga‐rich condition, provided by applying a very low V/III ratio, alters the growth rates of various planes of the defined nanostructure by increasing relative growth rate of the semi‐polar tilted m‐plane {1–101} that usually is the slowest growing plane under continuous growth conditions. This increased growth rate relative to the non‐polar m‐plane {1–100} and even the c‐plane (0001), permits the formation of GaN nanorods with nonpolar sidewalls. In addition, a new growth mode, called the NH₃‐pulsed mode, is introduced, utilizing the advantages of both the continuous mode and the lower growth rate pulsed mode to form nanorods. Finally, nanorods grown under the different growth modes are compared and discussed.     

Temperature-responsive culture surfaces for insect cell sheets to fabricate a bioactuator

ABSTRACT

For the first time, we have fabricated insect-derived cell sheets by using temperature-responsive culture surfaces having a phase-transition temperature below 25°C. We prepared the temperature-responsive cell culture surfaces (tissue culture polystyrene, TCPS) by grafting a copolymeric gel consisting of hydrophobic N-tert-butylacrylamide (tBAAm) and N-isopropylacrylamide (IPAAm) units. First, to characterize the hydrophilic and hydrophobic properties of the copolymeric gel-grafted surfaces, static water contact angles of each surface were measured at various temperatures. By increasing the amount of tBAAm in the grafted copolymeric gel, the transition temperature of the gel was shifted to lower temperatures. At 25°C, the grafted copolymeric gel was dehydrated, and the insect-derived cells (AeAl2 cells) adhered on all the copolymeric gel-grafted surfaces. At 20°C, AeAl2 cells cannot adhere on the P(IPAAm-1.62tBAAm)-TCPS surface (the initial molar ratio of IPAAm and tBAAm (tBAAm?=?1.62 mol%)) better than on other surfaces (TCPS and tBAAm?=?4.88, 8.17 mol%). These two findings implied that the lower critical solution temperature of the copolymeric gel-grafted-TCPS existed from 20°C to 25°C. The laminin-coated P(IPAAm-1.62tBAAm)-TCPS surface showed temperature-dependent cell attachment and detachment properties, while AeAl2 cells were not detached from the extracellular matrix uncoated P(IPAAm-1.62tBAAm)-TCPS surface. AeAl2 cells and insect muscle cells were harvested as the respective sheets.     

Pipe Heating System with Underground Water Tank for Snow Thawing and Ice Prevention on Roads and Bridge Decks

In order to prevent traffic accidents on snow-covered bridge decks, the writers developed a new pipe heating system that uses only groundwater stored in a large underground tank. The underground tank provides geothermal energy, i.e., groundwater of constant temperature, through heating pipes embedded in concrete pavements with no electric heater or fuel boiler. The pipe heating system was constructed at approximately 50% of the cost of comparable systems. In addition, there is a reduction of 10% in operating costs compared with the previous system. The present paper outlines its design and construction and provides fundamental data for the developed system. The piped heating system has kept the road conditions safe during the winter season by always removing the snow and ice from the heated road and bridge sections earlier than the surrounding roads. According to long-term temperature measurements, this system has prevented the road temperature from decreasing below 0°C, even through nighttime and morning periods. This report presents the system as an economical and effective solution for snow thawing and ice prevention.