Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa

We report here a novel type of ceramidase of Pseudomonas aeruginosa AN17 isolated from the skin of a patient with atopic dermatitis. The enzyme was purified 83,400-fold with an overall yield of 21.1% from a culture supernatant of strain AN17. After being stained with a silver staining solution, the purified enzyme showed a single protein band, and its molecular mass was estimated to be 70 kDa on SDS-polyacrylamide gel electrophoresis. The enzyme showed quite wide specificity for various ceramides, i.e. it hydrolyzed ceramides containing C12:0-C18:0 fatty acids and 7-nitrobenz-2-oxa-1, 3-diazole-labeled dodecanoic acid, and not only ceramide containing sphingosine (d18:1) or sphinganine (d18:0) but also phytosphingosine (t18:0) as the long-chain base. However, the enzyme did not hydrolyze galactosylceramide, sulfatide, GM1, or sphingomyelin, and thus was clearly distinguished from a Pseudomonas sphingolipid ceramide N-deacylase (Ito, M., Kurita, T., and Kita, K. (1995) J. Biol. Chem. 270, 24370-24374). This bacterial ceramidase had a pH optimum of 8.0-9.0, an apparent Km of 139 microM, and a Vmax of 5.3 micromol/min/mg using N-palmitoylsphingosine as the substrate. The enzyme appears to require Ca2+ for expression of the activity. Interestingly, the 70-kDa protein catalyzed a reversible reaction in which the N-acyl linkage of ceramide was either cleaved or synthesized. Our study demonstrated that ceramidase is widely distributed from bacteria to mammals.     

Bone Marrow-Mesenchymal Stromal Cell Secretome as Conditioned Medium Relieves Experimental Skeletal Muscle Damage Induced by Ex Vivo Eccentric Contraction

Bone marrow-mesenchymal stem/stromal cells (MSCs) may offer promise for skeletal muscle repair/regeneration. Growing evidence suggests that the mechanisms underpinning the beneficial effects of such cells in muscle tissue reside in their ability to secrete bioactive molecules (secretome) with multiple actions. Hence, we examined the effects of MSC secretome as conditioned medium (MSC-CM) on ex vivo murine extensor digitorum longus muscle injured by forced eccentric contraction (EC). By combining morphological (light and confocal laser scanning microscopies) and electrophysiological analyses we demonstrated the capability of MSC-CM to attenuate EC-induced tissue structural damages and sarcolemnic functional properties’ modifications. MSC-CM was effective in protecting myofibers from apoptosis, as suggested by a reduced expression of pro-apoptotic markers, cytochrome c and activated caspase-3, along with an increase in the expression of pro-survival AKT factor. Notably, MSC-CM also reduced the EC-induced tissue redistribution and extension of telocytes/CD34⁺ stromal cells, distinctive cells proposed to play a “nursing” role for the muscle resident myogenic satellite cells (SCs), regarded as the main players of regeneration. Moreover, it affected SC functionality likely contributing to replenishment of the SC reservoir. This study provides the necessary groundwork for further investigation of the effects of MSC secretome in the setting of skeletal muscle injury and regenerative medicine.     

Crystallographic Texture Development in Bismuth Sodium Titanate Prepared by Reactive-Templated Grain Growth Method

Bi_0.5Na_0.5TiO₃ (BNT) and 0.94Bi_0.5Na_0.5TiO₃·0.06BaTiO₃ (BNT–BT) bulk ceramics with extensive 〈100〉 texture were prepared by the reactive-templated grain growth method, using platelike Bi₄Ti₃O₁₂ (BIT) particles as templates for BNT. Calcined compacts were composed of matrix grains with random orientation and 〈100〉-oriented grains transformed from aligned BIT particles, and the texture developed by the growth of oriented grains during sintering. Ceramics with extensive texture were obtained by using the starting mixture containing the maximum concentration of platelike BIT to form the maximum volume fraction of oriented grains.     

A PRACTICAL METHOD FOR RECTIFYING INTERFACIAL TENSION DATA TO PREDICT THE SHAPE OF INTERFACES IN AN IMMISCIBLE LIQUID/LIQUID/GAS SYSTEM

The shape of interfaces formed by the contact of two liquid phases, immiscible with each other, and a gaseous phase can be predicted on the condition that the tension to work at each interface is known with a sufficient accuracy. In general, interfacial tension data available are not so accurate, however, as to be useful as they are in predicting the shape of interfaces, particularly when the spreading coefficient of either liquid (liquid 1) on the other (liquid 2) has a negative value of a small magnitude. Presented in this note is a simple method to rectify the interfacial tension data, with the aid of a measurement of the radius of a lens of liquid 1 of a known volume placed on the horizontal surface of liquid 2, and thereby make them usable for predicting interfacial geometries. The method is tested by applying it to benzene/water and n-pentane/water systems.     

A new maritime lifesaving multiple-effect solar still design

This paper presents a new maritime lifesaving multiple-effect solar still design where several extended wicks feed seawater to their evaporating areas by capillary force, and the resulting water vapor diffuses and condenses on facing wicks with the condensate flowing through the wicks into storage bags. A theoretical analysis showed that a temperature drop through diffusion layers between evaporating and condensing wicks slowly increases in the main evaporating areas and rapidly near salt depositions whereas the evaporation rate decreases slowly in the main area and rapidly near salt depositions. These changes are caused by increases in salt concentrations and resulting boiling point elevation. With steady-state transfer analysis, the proposed still is predicted to produce about 15 kg m ⁻²d⁻¹ fresh water on a sunny day of 22 MJm⁻²d⁻¹ solar radiation, showing a potential to be a maritime lifesaving desalinator.     

Changes in the Peeled Surfaces of Copper Thin Films Deposited on Polyimide Substrates After Heat Treatment

Peel strength between a copper (Cu) thin film and a polyimide (pyromellitic dianhydride-oxydianiline, or PMDA-ODA) substrate is reduced by heat treatment at 150°C in air. In this work, we investigated the peel strength, the morphology of the interface between Cu films and polyimide substrates using optical microscopy and electron microscopy, and chemical change of the interface using Auger electron spectroscopy (AES) and micro X-ray photoelectron spectroscopy (XPS). The analysis showed that CuO “lumps” were present on the peeled surface of PMDA-ODA after heat treatment at 150°C in air. The peeled surfaces of other polyimide substrates were also analyzed: biphenyl dianhydride-para phenylene diamine (BPDA-PDA) and biphenyl dianhydride-oxydianiline (BPDA-ODA). CuO lumps were present on the peeled surface of BPDA-ODA after the heat treatment, but not that of BPDA-PDA. Compared with the adhesion strength for the Cu thin film, the adhesion strength was high for the Cu/PMDA-ODA and Cu/BPDA-ODA laminates, but the adhesion strength was very low for the Cu/BPDA-PDA laminate. This low strength is the reason that CuO lumps were not detected on the peeled surface of the BPDA-PDA substrate. These CuO lumps were related to the adhesion degradation of the Cu/polyimide laminates after the heat treatment.     

Probability of precision micro-machining of insulating Si3N4 ceramics by EDM

Electrical discharge machining (EDM) is used as a precision machining method for the electrically conductive hard materials with a soft electrode material. But recently we succeeded to machine on insulating material by EDM. The technology is named as an assisting electrode method. The EDMed surface is covered with the electrical conductive layer during discharge. The layer holds the electrical conductivity during discharge. For micro-EDM, the wear of tool electrode becomes lager ratio than the normal machining. So the micro-machining is extremely difficult to get the precision sample.

In this paper to obtain a fine and precise ceramics sample, some trials were carried out considering the EDM conditions, tool electrodes material and assisting electrode materials. Insulating Si₃N₄ ceramics were used for workpiece. The machining properties were estimated by the removal rate and tool wear ratio. To confirm the change of micro-machining process, the discharge waveforms were observed. The micro-machining of the Ø0.05 mm hole could be machined with the commercial sinking electrical discharge machine.     

Developing a Higher-Cycled Product Design CAE Model： The Evolution of Automotive Product Design and CAE

To help corporations survive amidst worldwide quality competition, the authors have focused on the strategic development of a higher-cycled product design CAE model employing a highly reliable CAE analysis technology component model. Their efforts are part of principle-based research aimed at evolving product design and CAE development processes to ensure better quality assurance. To satisfy the requirements of developing and producing high quality products while also reducing costs and shortening development times, the effectiveness of this model was verified by successfully applying it to the technological problems of loosening bolts and other product design bottlenecks at auto manufacturers.     

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.     

Stick-slip motion control based on cutter location data for an orthogonal-type robot

In this article, a new desktop orthogonal-type robot, which has the capacity of stick-slip motion control based on cutter location data, is presented for lapping small metallic molds with a curved surface. The robot consists of three single-axis devices with a high position resolution of 1 μm. A thin wooden stick tool with a ball-end shape is attached to the tip of the z-axis. In order to improve the lapping performance, a novel stick-slip motion control method is developed in the control system. The small stick-slip motion is orthogonally generated in the direction of the tool’s movement. The effectiveness of stick-slip motion control is examined through an actual lapping test of an LED lens cavity.