Properties and structure of spermidine acetyltransferase in Escherichia coli

Spermidine acetyltransferase (SAT) from Escherichia coli was purified about 40,000-fold. The molecular mass of native SAT was 95 kDa, and it consisted of four identical subunits. The products formed from the reaction of acetyl-CoA with spermidine by SAT were N1- and N8-acetylspermidine. The Km values for acetyl-CoA, spermidine, and spermine were 2 microM, 1.29 mM, and 220 microM, respectively. The enzymatic activity increased by 2.5-3.5-fold under the condition of poor nutrition but not in response to cold shock or high pH. By using synthetic oliogonucleotides deduced from amino acid sequences of the peptides in SAT, a polymerase chain reaction product with a length of 250 nucleotides was obtained. Using this polymerase chain reaction product, the gene encoding SAT (speG) was cloned and mapped at 35.6 min in the E. coli chromosome. E. coli cells transformed with the cloned speG gene increased SAT activity by 8-40-fold. The gene encoded a 186-amino acid protein, but SAT consisted of 185 amino acids because the initiator methionine was liberated from the protein. Thus, the predicted molecular mass was 21,756 Da. Significant similarity to aminoglycoside acetyltransferase and peptide N-acetyltransferase was observed in the amino acid sequence 87-141, and some similarity with spermidine-preferential binding protein (potD protein) in the spermidine-preferential uptake system was observed in the amino acid sequence 122-141. The results suggest that the active center of SAT may be located in the COOH-terminal portion.     

The relationship between electropermeabilization and cell cycle and cell size of Saccharomyces cerevisiae

Yeast cells (Sacchromyces cerevisiae) in 0.9% NaCl solution containing phloxine B (dye) were treated by an application of a rectangular electric pulse. We input microscopic images of the yeast suspensions after the application into a computer, and measured whether each cell dyes or not, the phase in the cell cycle, and each cell size, using the software we had developed. After those measurements, we discussed the relationship between the yield of electropermeabilization (the ratio of dyed cells to the total cell number) and the phase in the cell cycle, and cell size. From the results, it was found that the yeast cells from S-phase to M-phase (S-M phase) in the cell cycle tend to be more permeated than G1-phase yeast cells, and in both phases the yield decreases with the increase in cell size.     

Porcine enamel matrix derivative enhances the formation of reparative dentine and dentine bridges during wound healing of amputated rat molars

We examined the biological effects of porcine enamel matrix derivative (EMD; Emdogain) on the formation of reparative dentine and dentine bridges in rat molars after pulp amputation. The pulp chambers of upper molars of Wistar rats were perforated and the amputated pulp surfaces were directly capped with either EMD or its carrier propylene glycol alginate (PGA) as control. The cavities were then restored with glass-ionomer cement. On post-amputation days 4-30, the dissected maxillae were examined by light and electron microscopy. In PGA-capped pulp, reparative dentine had been formed over the dentine walls under the prepared cavity on day 7 post-amputation and its thickness extended until day 30. On day 30, as well as reparative dentine formation, diffuse calcification had occurred beneath the amputated wound surfaces. Dentine bridge formation under the amputated coronal pulp surface was observed in 18.2% of amputated pulp on day 30. In EMD-capped pulp, reparative dentine had already been formed by odontoblast-like cells over the dentine walls, already on day 4 post-amputation, and its thickness extended until day 30. The Ca and P weight % and Ca/P ratio of reparative dentine matrix were similar to those of pre-existing dentine matrix, and these values were not different between PGA and EMD-capped pulp. Dentine bridge formation was observed in 27.3% of EMD-capped pulp on day 30. Our results suggest that EMD enhances the formation of both reparative dentine and dentine bridges during wound healing of amputated rat molar pulp.     

A novel anionic surfactant templating route for synthesizing mesoporous silica with unique structure

Anionic surfactants are used in greater volume than any other surfactants because of their highly potent detergency and low cost of manufacture. However, they have not been used as templates for synthesizing mesoporous silica. Here we show a templating route for preparing mesoporous silicas based on self-assembly of anionic surfactants and inorganic precursors. We use aminosilane or quaternized aminosilane as co-structure-directing agent (CSDA), which is different from previous pathways. The alkoxysilane site of CSDA is co-condensed with inorganic precursors; the ammonium site of CSDA, attached to silicon atoms incorporated into the wall, electrostatically interacts with the anionic surfactants to produce well-ordered anionic-surfactant-templated mesoporous silicas (AMS). These have new structures with periodic modulations as well as two-dimensional hexagonal and lamellar phases. The periodic modulations may be caused by the coexistence of micelles that differ in size or curvature, possibly owing to local chirality. These mesoporous silicas provide a new family of mesoporous materials as well as shedding light on the structural behaviour of anionic surfactants.     

A novel strategy of cell targeting based on tissue-specific expression of the ecotropic retrovirus receptor gene

Gene transfer into specific tissues or cell types is a key technique in the development of gene therapy. Modification of vector particles such that they selectively bind to the target cells has been attempted, but the limitation of this approach is the low transduction efficiency. Here, we show that a two-step gene transfer system can be used for efficient cell targeting. With this strategy, and using a high-titer adenoviral vector containing a tissue-specific promoter, we have engineered a system in which only target cells become susceptible to retrovirus-mediated transduction. In a model experiment, we constructed an adenoviral vector (Ad.AFPEcoRec) containing the ecotropic retrovirus receptor (EcoRec) gene under the control of the alpha-fetoprotein (AFP) promoter. A binding assay showed that after transduction with AD.AFPEcoRec, EcoRec molecules were efficiently expressed in AFP+HepG2 cells, but not in AFP-HeLa and AFP-HLE cells. The EcoRec-expressing HepG2 cells could be stably transduced with ecotropic retroviral vectors, whereas HeLa and HLE cells remained highly resistant to retrovirus-mediated gene transfer. The apparent titer on HepG2 cells was greater than 2 x 10(5) CFU/ml. Because various tissue-specific promoter/enhancer elements are available, the two-step system could be used as a general strategy for both ex vivo and in vivo targeted gene transfer.     

Nonsalt 1‐(arylmethyloxy)pyrene photoinitiators capable of initiating cationic polymerization

Analysis of photoproducts derived from 1‐(methoxynaphthalen‐1‐ylmethyloxy)pyrene initiators and polymer end groups demonstrated that methoxynaphthalen‐1‐ylmethyl carbocation is involved in the initiation steps for both styrene (St) and cyclohexene oxide (CHO) polymerization. Charge transfer from the pyrenyloxy oxygen atom to the methoxynaphthalen‐1‐ylmethyl chromophore in the singlet excited state is assumed to be responsible for the efficient generation of the carbocation species, which also initiates the copolymerization of St and CHO. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40510.     

Survey of actual viewing conditions at home and appropriate luminance of LCD‐TV screens

Abstract— To understand actual viewing conditions at home is important for TV design. And the preferred luminance level of LCD TVs under actual viewing conditions is also important in order to obtain both good picture quality and low power consumption. The actual viewing conditions of households and the preferred luminance levels was investigated. In a field test of 83 households, the display luminance, screen illuminance, and viewing locations were measured on site. In laboratory experiments, young and elderly subjects adjusted the luminance of an LCD‐TV screen to their preferred levels under different screen illuminance levels, angular screen sizes, and average luminance levels (ALL) of the images. As a result, two equations, which represent the preferred luminance level of LCD‐TV screens corresponding to different viewing conditions for young and elderly subjects were obtained. When the ALL of the images was 25% and the screen illuminance and angular screen size were set at 100 lx and 20°, respectively, the preferred luminance was 1 60 cd/m² for the young subjects and 248 cd/m² for the elderly subjects. By using the setting of the preferred luminance of an LCD TV under actual viewing conditions, it is possible to conserve energy consumption.     

Magnesium interlayered diamond coating on silicon

Diamond thin films have been deposited using hot filament chemical vapour deposition technique on manually scratched p-Si(1 0 0) substrate, with and without magnesium interlayer. In spite of magnesium melting point being lower (T_m = 649 °C) than the growth temperature of the substrate (T_s  750 °C) used in these experiments, it was found that high quality diamond films could be grown on Mg covered substrate. A liquid substrate is probably generated during the diamond film growth. Raman spectroscopy analysis exhibited only the triply degenerate, zone centre optical phonon peak at 1333 cm⁻¹ indicating that nearly stress free crystallites were present. Broadening of the Raman peak (11.76 cm⁻¹) indicates that some small crystallites also are present. Scanning electron and atomic force microscopy accompanied by X-ray diffraction analysis where used to compare the details of diamond film growth directly on scratched Si(1 0 0) and Mg interlayered scratched Si(1 0 0) substrates.