Chromosomal DNA patterns and gene stability of Pichia pastoris

We have clearly resolved four chromosomal bands from four Pichia pastoris (Komagataella pastoris) strains by using contour-clamped homogeneous electric field gel electrophoresis. The size of the P. pastoris chromosomal bands ranged from 1.7 Mb to 3.5 Mb and total genome size was estimated to be 9.5 Mb to 9.8 Mb; however, chromosome-length polymorphisms existed among four strains. Thirteen cloned genes isolated from strain GTS115 were assigned to the separated chromosomes, revealing that different hybridization patterns were observed in the AOX2 and URA3 genes among strains. P. pastoris is frequently used as an efficient host for heterologous gene expressions. We analysed chromosomal stability of strain GTS115-derived recombinant cell expressing human serum albumin during serial cultivation under the condition of vegetative and non-selective growth. No chromosomal rearrangements were observed and the expression constructs integrated into the his4 locus on chromosome I were very stable even at 83 generations, suggesting that stable expression would be carried out even in large-scale fermentation.     

A novel method of detecting Staphylococcus aureus heterogeneously resistant to vancomycin (hetero-VRSA)

We have developed a new method of detecting clinical S. aureus strains possessing heterogeneous resistance to vancomycin (hetero-VRSA). The method exploits characteristic antagonism between vancomycin and beta-lactam antibiotics against the strain Mu3, a representative hetero-VRSA. The method comprises of the following procedures: 1) overnight culture of bacteria is streaked on brain heart infusion agar containing 4 micrograms/ml of vancomycin, 2) paper discs impregnated with any one of beta-lactam antibiotics are placed onto the plate, and 3) the plate is incubated at 37 degrees C and the cell growth was observed after 24 h and 48 h of incubation. The Mu3 cells were grown only around the beta-lactam discs due to the antagonism between vancomycin and beta-lactam against Mu3 cells. A total of 321 MRSA clinical strains isolated in 1995 and 1996 from 12 medical facilities in Japan were tested with this method. A total of 39 strains (12.1%; 24 h) and 67 strains (20.96%; 48 h) grew around the beta-lactam discs. All the 10 strains (representing 10 facilities), tested with the analysis of resistant subpopulations to vancomycin, showed similar heterogeneous patterns as observed with Mu3. The method was considered as a convenient and rapid substitute for the population analysis, the authentic method for the detection of hetero-VRSA strains.     

The relationship between cardiac contraction and intramyocardialhemodynamics

Since coronary venous flow is squeezed out from the myocardial vascular bed by the extravascular compressive force of heart muscle, the coronary venous system provides a suitable model to investigate the relationship between cardiac contraction and intramyocardial hemodynamics. To investigate coronary venous flow velocity in more detail, we previously developed a laser Doppler velocimeter (LDV) with an optical fiber. Furthermore, we have recently developed a needle probe charge-coupled device (CCD) videomicroscope to visualize intramyocardial microvessels. In this article, we report I) blood flow velocity waveforms in the epicardial small veins of the left ventricle and 2) subendocardial venular diameter changes during a cardiac cycle     

Study of asynchronizing phenomena using generator model with field mutual leakage reactance

This paper describes the effect of mutual leakage reactance in the rotor circuit on the analysis of generator transient characteristics. The authors derive a detailed equivalent circuit model for a 48P‐15,750‐kVA generator, analyze sudden short circuits and asynchronization phenomena of the generator by simulations with an electromagnetic transients program (ATP‐EMTP), and compare the results with experimental data. © 2005 Wiley Periodicals, Inc. Electr Eng Jpn, 153(3): 31–40, 2005; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.20147     

Turkey rhinotracheitis virus isolated from broiler chicken with swollen head syndrome in Japan

Turkey rhinotracheitis (TRT) virus was first isolated from a commercial broiler chicken with swollen head syndrome (SHS) in Japan. At the same time, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), avian reovirus (ARV), Escherichia coli (E.coli), Morganella morganii, and Proteus mirabilis were also isolated from the same broiler chicken. The presence of antibodies to TRT virus was confirmed in the sera of 34-day-old chickens of the flock with SHS, however the antibodies to TRT virus were undetectable in the sera of 17-day-old chickens. In this investigation, we confirmed avian pneumovirus infection in chickens in Japan, and the virus and other agents may be considered as a cause of SHS.     

Melting and glass transition of radiation-induced graft polyethylene

Melting and glass transition data are reported employing DSC for styrene-grafted high-density polyethylene obtained by γ radiation. Judging from the data of the melting point and the heat of fusion, the grafted polystyrene had no effect on the polyethylene crystallites, but the half-width of the thermogram was observed to increase slightly, showing an effect on the crystallite size distribution. As no effect was observed on the glass transition temperature by grafting, the amorphous region of the polyethylene apparently was not affected. It is suggested, therefore, that the free volume or segmental mobility will not be decreased by radiation-induced grafting. Very few but long grafted chains had negligible effect on the average polyethylene chain length available for segmental motion, and grafted polystyrene should be expected to differ little from the styrene homopolymer in thermal motions.     

Effects of grain boundary on dielectric properties in fine-grained BaTiO3 ceramics

BaTiO₃ dielectric ceramics, which have no core–shell structure, were prepared. High-resolution transmission electron microscope (TEM) and the newly developed chemical etching analysis revealed the composition of the grain boundary. The grain boundary has an important role in stabilizing temperature dependence of a dielectric constant. The grain boundary causes a large stress against grains, resulting in a change of the domain structure. So, the stress from the grain boundary is considered to depress the dielectric abnormality at the Curie temperature.     

Adsorption of oxygen in an activated europium ion-exchanged mordenite

The adsorption of oxygen in an activated europium ion-exchanged mordenite(Eu-M) was studied over the temperature range 25–600°C by the measurement of fluorescence of Eu₂₊ ion and a temperature programmed desorption (TPD) spectra of oxygen. When oxygen was exposed to a activated Eu-M, the intensity of emission band for Eu²⁺ ion extremely decreased. After the adsorption of oxygen at room temperature, the emission intensity was increased with a rise of degassing temperature and restored to the original emission intensity above 300°C. While, in Eu-M, at least four different states of adsorbed oxygen were indicated by the appearance of four TPD peaks with peak maxima located at about 70°C(α), 220°C(β), 300°C(γ) and >500°C(δ). The intensity of TPD peaks was dependent on the adsorption temperature. In the case of adsorption at 300°C or 600°C, the total amount of desorbed oxygen corresponded to one oxygen molecule adsorbing per Eu²⁺ ion.