Functional Properties of the Catalytic Domain of Mouse Acidic Mammalian Chitinase Expressed in Escherichia coli

Mouse acidic mammalian chitinase (AMCase) plays important physiological roles in defense and nutrition. AMCase is composed of an N-terminal catalytic domain (CatD) and a C-terminal chitin-binding domain (CBD). We expressed CatD of mouse AMCase as a recombinant fusion protein with Protein A and V5-His in Escherichia coli (Protein A-CatD-V5-His), evaluated its functional properties and compared them to the full-length AMCase (Protein A-AMCase-V5-His). Under our experimental conditions, the chitinolytic activity of both proteins against 4-nitrophenyl N,N''-diacetyl-β-d-chitobioside was equivalent with regard to their specific enzymatic activities, optimal pH and temperature as well as to the pH and temperature stability. CatD bound to chitin beads and cleaved the N-acetylglucosamine hexamer, colloidal and crystalline chitin as well as the shrimp shell, and released primarily N,N''-diacetylchitobiose fragments at pH 2.0. These results indicate that the primary structure of CatD is sufficient to form a proper tertiary structure required for chitinolytic activity, recognize chitin substrates and degrade them in the absence of a CBD. Our recombinant proteins can be used for further studies evaluating pathophysiological roles of AMCase in different diseases.     

Difference in degrading p-nitrophenol between indigenous bacteria in a reactor

p-Nitrophenol (PNP) -degrading bacteria were isolated from a reactor using a mineral salt medium containing a low and high PNP concentration. We isolated two bacterial species, Pseudomonas sp. YTK17 and Rhodococcus opacus YTK32, that utilize PNP as their sole source of carbon and energy. These strains exhibited differences in PNP degradation activity in relation to PNP concentration. Strain YTK17 showed a high level of degradation following pre-exposure to a low PNP concentration, whereas strain YTK32 required a relatively high PNP concentration for degradation to occur. These results indicated that phylogenetically and physiologically different types of PNP-degrading bacteria coexisted in a reactor.     

Nonsense-mutated inlA and prfA not widely distributed in Listeria monocytogenes isolates from ready-to-eat seafood products in Japan

InlA is a surface protein participating in the entry of Listeria monocytogenes into mammalian non-phagocytic cells. PrfA is a positive regulatory factor that regulates the expression of a set of virulence genes. Recent studies revealed that some L. monocytogenes strains have a truncated form of these proteins because of nonsense mutations in their sequences, and these truncations contribute to the significant reduction in virulence of this pathogen. In this study, sequence analyses of inlA and prfA among L. monocytogenes isolated from ready-to-eat seafood revealed that only one out of 59 isolates had a nonsense-mutated inlA and all had non-mutated prfA. This indicated that these strains could be fully virulent based on the sizes of these proteins.     

食用香甜味沙拉酱对β胡萝卜素吸收的影响

目的:通过动物实验、体外实验确认食用香甜味沙拉酱对β-胡萝卜素吸收的影响效果。方法:采用高效液相色谱(high performance liquid chromatography,HPLC)技术进行β-胡萝卜素定量分析;参考93号饲料(AIN-93)配方,分组连续喂养大鼠12 d,测定不同组别大鼠肝脏中β-胡萝卜素的含量;配制人工胃液,分别将磨碎的芒果与香甜味沙拉酱和大豆油混合,模拟胃液蠕动搅拌不同时间,并测定油相中β-胡萝卜素含量;分别给大鼠灌胃等量的β-胡萝卜素以及油量相等的香甜味沙拉酱与大豆油,比较香甜味沙拉酱与大豆油在大鼠胃中停留的时间并测定油相中β-胡萝卜素含量。结果:大鼠肝脏中β-胡萝卜素含量为香甜味沙拉酱组大豆油组(P0.05);体外模拟实验发现,芒果更容易分散于香甜味沙拉酱体系,油相中β-胡萝卜素含量为香甜味沙拉酱组大豆油组,搅拌60 min组搅拌30 min组(P0.05);大鼠在灌胃30 min后胃液中β-胡萝卜素含量为香甜味沙拉酱组大豆油组(P0.05),且香甜味沙拉酱在胃中停留的时间更长。结论:食用香甜味沙拉酱比单纯食用大豆油更能够促进β-胡萝卜素的吸收转化;香甜味沙拉酱更能促进富含β-胡萝卜素水果中的β-胡萝卜素的吸收。     

Heat transfer control in an enclosure by using a rotating plate and stratified fluids

The effects of an inner rotating plate with horizontal axis on the heat transfer between two vertical walls in a rectangular enclosure with stratified fluid layers were investigated experimentally. The aspect ratio of the enclosure height/width was 1 throughout the experiments. An acrylic plate with small thermal conductivity was installed horizontally at the center of the square enclosure, and was rotated at various speeds for normal and reverse rotations by using the motor installed outside of the enclosure. Purified water and silicon oil were used as the working fluids and were stratified in the enclosure. It is shown here that the heat transfer rate of the enclosure with stratified fluid layers differs largely from that of the enclosure with a single fluid layer. Namely, the heat transfer coefficient increases rapidly at a low rotating speed range, and maintains an almost constant value at a high rotating speed range. © 2003 Wiley Periodicals, Inc. Heat Trans Asian Res, 32(6): 489–500, 2003; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/htj.10081     

Fluid flow and heat transfer of natural convection adjacent to upward‐facing inclined heated plates

Turbulent transition mechanism and local heat transfer characteristics of the natural convective flows over upward‐facing inclined plates were investigated experimentally. The experiments were performed in the range of modified local Rayleigh numbers from 10⁴ to 8 × 10¹⁴ and of inclination angles θ from 0 to 90°. The flow fields over the plate and the surface temperatures of the plate were visualized with dye and liquid crystal thermometry. The results showed that longitudinal vortices play a main role in the turbulent transition over the plate of θ < 72°. These vortices appear first in the laminar boundary layer, then detach from the plate and, finally become distorted. It is found that the heat transfer is enhanced markedly by the detachment and the distortion of these vortices. The local heat transfer coefficients were measured in the laminar, transitional, and turbulent regions. The results show that the coefficients in the turbulent region become identical and independent of inclination angles. © 2003 Wiley Periodicals, Inc. Heat Trans Asian Res, 32(3): 278–291, 2003; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/htj.10091     

Solidification in a water‐saturated porous medium when convection is present (response of solid‐liquid interface due to time‐varying cooling temperature)

A fundamental study on solidifying phenomenon in a rectangular space filled with water‐saturated porous medium has been carried out with a system, cooled from the upper boundary and heated from below, where vigorous convection develops in the un‐solidified liquid layer. The dynamic response of the solid‐liquid interface to the periodical cooling temperature with the bottom boundary kept at constant temperature T_H = 20°C, is investigated experimentally. In particular, the amplitude of the interface and the phase lag in respect to the oscillating cooling temperature have been monitored for various periods and average temperatures. A one‐dimensional numerical model, based on an assumption of constant heat flux from the vigorously convecting liquid regime has been also developed. The numerical model predicts quite well the time‐dependent behavior of the horizontally averaged ice‐layer thickness observed in the experiments. Our general findings are that the amplitude increases proportionally to the temperature fluctuation period and that both the thicker solid layer and the shorter period cause greater phase lags. © 2006 Wiley Periodicals, Inc. Heat Trans Asian Res, 35(4): 294–308, 2006; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/htj.20109     

Heat transfer regulation in a rectangular enclosure using a rotating plate

The effects of an inner rotating plate with horizontal axis on the heat transfer in a differentially heated vertical enclosure were investigated experimentally. The aspect ratio of the enclosure height/width was 1 throughout the experiments. An acrylic plate with a small thermal conductivity was installed horizontally at the center of the square enclosure, and was rotated at various speeds for normal and reverse rotations by using the motor attached outside of the enclosure. Purified water was used as the working fluid. The flow pattern was sketched by a visualization experiment using aluminum powder. The heat transfer results were also compared with those from a previous paper on a rotating cylinder. It is clarified here that the heat transfer rate of the enclosure depends largely on the parameter Gr_w/Re_ω², and is characterized by three regions. The heat transfer rate of the enclosure with a rotating plate is somewhat larger than that of a rotating cylinder in the forced convection region. The rotating plate used here will be useful for regulation of wide‐ranging heat transfer. © 2003 Wiley Periodicals, Inc. Heat Trans Asian Res, 32(4): 342–353, 2003; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/htj.10099     

IFMIF specifications from the users point of view

This paper summarizes the proposals and findings of the IFMIF Specification Working Group established to update the users requirements and top level specifications for the facility. Special attention is given to the different roadmaps of fusion pathway towards power plants, of materials R&D and of facilities and their interactions. The materials development and validation activities on structural materials, blanket functional materials and non-metallic materials are analyzed and specific objectives and requirements to be implemented in IFMIF are proposed. Emphasis is made in additional potential validation activities that can be developed in IFMIF for ITER TBM qualification as well as for DEMO-oriented mock-up testing.