Poly[(R)-3-hydroxybutyrate] formation in Escherichia coli from glucose through an enoyl-CoA hydratase-mediated pathway

In this study, a new metabolic pathway for the synthesis of poly[(R)-3-hydroxybutyrate] [P(3HB)] was constructed in a recombinant Escherichia coli strain that utilized forward and reverse reactions catalyzed by two substrate-specific enoyl-CoA hydratases, R-hydratase (PhaJ) and S-hydratase (FadB), to epimerize (S)-3HB-CoA to (R)-3HB-CoA via a crotonyl-CoA intermediate. The R-hydratase gene (phaJ(Ac)) from Aeromonas caviae was coexpressed with the PHA synthase gene (phaC(Re)) and 3-ketothiolase gene (phaA(Re)) from Ralstonia eutropha in fadR mutant E. coli strains (CAG18497 and LS5218), which had constitutive levels of the beta-oxidation multienzyme FadB(Ec). When grown on glucose as the sole carbon source, the cells accumulated P(3HB) up to an amount 6.5 wt% of the dry cell weight, whereas the control cells without phaJ(Ac) or fadR mutation accumulated significantly smaller amounts of P(3HB). These results suggest that PhaJ(Ac) and FadB(Ec) played an important role in supplying monomers for P(3HB) synthesis in the pathway. Furthermore, by using this pathway, a P(3HB)-concentration-dependent fluorescent staining screening technique was developed to rapidly identify cells that possess active R-hydratase.     

Syntheses of alpha-arbutin-alpha-glycosides and their inhibitory effects on human tyrosinase

Alpha-arbutin is a tyrosinase inhibitor. We synthesized alpha-arbutin-alpha-glycosides by the transglycosylation reaction of cyclomaltodextrin glucanotransferase from Bacillus macerans using alpha-arbutin and starch as acceptor and donor molecules, respectively. We isolated and characterized two major products from the reaction mixture. The structural analyses using 13C- and 1H-NMR spectroscopy proved that they were 4-hydroxyphenyl alpha-maltoside (alpha-Ab-alpha-G1) and 4-hydroxyphenyl alpha-maltotrioside (alpha-Ab-alpha-G2). Both alpha-Ab-alpha-G1 and alpha-Ab-alpha-G2 exhibited competitive-type inhibition on human tyrosinase as alpha-arbutin does. Their K(i) values were calculated to be 0.6 mM and 2.8 mM, respectively, which is slightly and significantly higher than that of alpha-arbutin (0.2 mM).     

Effects of Fruiting Bodies of Basidiomycetes on Yeast Growth Rates

The effects of edible fruiting of Basidiomycetes on growth of Saccharomyces cerevisiae and other yeast strains were examined. The growth rates were significantly increased in the presence of fruiting bodies but there was no significant difference growth yield between cultures with and without fruiting bodies. Growth rates of yeast cells were promoted in both synthetic and natural media.     

The tomato DWARF enzyme catalyses C-6 oxidation in brassinosteroid biosynthesis

Brassinosteroids (BRs) are steroidal plant hormones essential for normal plant growth and development. Mutants in the biosynthesis or perception of BRs are usually dwarf. The tomato Dwarf gene (D), which was predicted to encode a cytochrome P450 enzyme (P450) with homology to other P450s involved in BR biosynthesis, was cloned previously. Here, we show that DWARF catalyses the C-6 oxidation of 6-deoxocastasterone (6-deoxoCS) to castasterone (CS), the immediate precursor of brassinolide. To do this, we first confirmed that the D cDNA complemented the mutant light- and dark-grown phenotypes of the extreme dwarf (dx) allele of tomato. To identify a substrate for the DWARF enzyme, exogenous application of BR intermediates to dx plants was carried out. C-6 oxoBR intermediates enhanced hypocotyl elongation whereas the C-6 deoxoBR, 6-deoxoCS, had little effect. Quantitative analysis of endogenous BR levels in tomato showed mainly the presence of 6-deoxoBRs. Furthermore, dx plants were found to lack CS and had a high level of 6-deoxoCS in comparison to D plants that had CS and a lower level of 6-deoxoCS. Confirmation that DWARF catalyzed the C-6 oxidation of 6-deoxoCS to CS was obtained by functional expression of DWARF in yeast. In these experiments, the intermediate 6alpha-hydroxycastasterone was identified, indicating that DWARF catalyzes two steps in BR biosynthesis. These data show that DWARF is involved in the C-6 oxidation in BR biosynthesis.     

Visible Light Irradiation of [60]Fullerene Causes Killing and Initiation of Transformation in Balb/3T3 Cells

Cell transformation in vitro is a model of carcinogenesis in vivo. Two-stage transformation assay increases the sensitivity of cells to chemicals and permits detection of carcinogens acting as initiating agents. [60]Fullerene (C₆₀) was cytotoxic in BALB/3T3 cells when it was irradiated by visible light, but not without light irradiation. Under conditions when C₆₀ was cytotoxic, it acted as an initiating agent for cell transformation, but it did not act as a complete transforming agent. the initiating activity of visible-light-irradiated C₆₀ was statistically significant in a modified two-stage transformation assay including a procedure for replating cells treated by C₆₀ and light, but it was equivocal in the standard two-stage transformation assay.     

Heat transfer characteristics of a reservoir embedded loop heat pipe (Heat transfer characteristics of a deployable radiator for use on the ETS‐VIII satellite while in orbit)

As heat generation in satellites increases, ensuring that they are provided with sufficient radiator panel area is an important problem. Deployable radiators, with radiator panels that are deployed post‐launch in space to increase the satellite's effective radiator panel area of the satellite, are becoming an important thermal control technology. A reservoir embedded loop heat pipe (RELHP) is used in deployable radiators as a heat transport device. A deployable radiator of this type was mounted on the ETS‐VIII satellite, which was launched on December 18, 2006 and injected into a geostationary orbit. The satellite is still operating without any significant issues over two years later. This paper investigates the heat transport characteristics of an RELHP system used in a deployable radiator in a geostationary orbital environment. This system can be successfully started up in a micro gravity environment. We also found that the sub‐cooling region is shorter in a micro gravity environment than in a terrestrial gravity environment, because there is less heat leakage into the reservoir in a micro gravity environment. © 2011 Wiley Periodicals, Inc. Heat Trans Asian Res; Published online in Wiley Online Library ( wileyonlinelibrary.com/journal/htj ). DOI 10.1002/htj.20346     

Design and control of a microrobotic system using magneticlevitation

Presents a prototype microrobot based on magnetic principles. Miniature items are to be transported and assembled in hazardous environments. A microrobot can be remotely operated with 3 DOF in an enclosed environment by transferring magnetic energy and optical signals from outside. The magnetic drive unit consists of 8 electromagnets (4 pairs), 2 permanent magnets, a return yoke and a pole piece. The microrobot is manipulated under the pole piece by regulating magnetic field. It consists of a magnetic head, a body (electronic circuit and batteries), and copper alloy ribbon ringers. A shape memory alloy actuator activates the fingers by illuminating/extinguishing several LED. PID controls were applied. To cope with uncertainties and variations in payload masses, an adaptive control law was also employed for positioning along the z axis to enable the controller parameters to be adjusted in real-time. Effectiveness of the control was verified by the results of several experiments. The microrobot has a net mass of 8.1 g and it can elevate and manipulate objects with masses up to 1.5 g within a volume of 29×29×26 mm³ with a precision of 0.05 mm     

Effect of aluminium on the polymerization silicic acid in aqueous solution and the deposition of silica of silica

The effect of aluminium on the polymerization of silicic acid was studied at pH 7, 8 and 9 in the aluminium concentration range of 0–26 ppm (Al) by spectrophotometry, gel chromatography and ²⁷Al NMR. Retarding and accelerating effects of aluminium on the growth of polysilicic acid particles and on the reaction between monosilicic acid and polysilicic acid were observed by changing the pH. It is suggested that the accelerating effect on the reaction between polysilicic acid particles is due to the formation of aluminium hydroxide on the surface of polysilicic acid. The rate of decrease in the monosilicic acid concentration in the presence of aluminium was faster than that in the absence of aluminium at pH 9, because monosilicic acid could be adsorbed rapidly on the aluminium hydroxide. From the results it was presumed that the formation of aluminium hydroxide on the solid surface may accelerate the deposition of silicic acid from geothermal water.     

Possible relation of osteopontin to development of psammoma bodies in human papillary thyroid cancer

Fine-needle aspiration cytology (FNAC) is the most useful procedure for the evaluation of thyroid nodules. The requirement for repeated aspirations in the follow-up of benign nodular thyroid disease, however, is controversial. To determine the value of re-aspirations in benign nodular thyroid disease, we studied 457 fine-needle reaspirations performed on 216 patients (197 female, 19 male) aged 42.9+/-12 years with uninodular (n = 65) and multinodular (n = 151) thyroid disease. Two hundred fifty-seven of these were second, 137 were third, 46 were fourth, and 17 were fifth re-aspirations of the same nodule, performed in a mean follow-up time of 43.9+/-31 (3-156) months. FNAC results were benign in 407 (89%), insufficient for diagnosis in 31 (6.8%), suspicious in 16 (3.5%), and papillary carcinoma (PC) in 3 (0.7%). An initial benign diagnosis did not change after multiple aspirations in 213 (98.61%) of the cases. Three patients with initial aspirations read as benign had a diagnosis of PC from their second biopsies, (diagnosis confirmed at surgery). Re-examination of the initial FNAC revealed atypical features in 1 of the 3 patients. These 3 patients likely represent a false-negative result of the initial FNAC rather than benign nodular disease transformed to a malignant one during the follow-up period. In conclusion, a second aspiration of clinically suspicious nodules may correct a few initial false-negative results, but routine additional re-aspirations are not useful for clinically stable disease.