Production of FAME from acid oil model using immobilized <Emphasis Type="Italic">Candida antarctica</Emphasis> lipase

Acid oil is a by-product in the neutralization step of vegetable oil refining and is an alternative source of biodiesel fuel. A model substrate of acid oil, which is composed of TAG and FFA, was used in experiments on the conversion to FAME by immobilized Candida antarctica lipase. FFA in the mixture of TAG/FFA were efficiently esterified with methanol (MeOH), but the water generated by the esterification significantly inhibited methanolysis of TAG. We thus attempted to convert a mixture of TAG/FFA to FAME by a two-step process comprising methyl esterification of FFA and methanolysis of TAG by immobilized C. antarctica lipase. The first reaction was conducted at 30°C in a mixture of TAG/FFA (1∶1, wt/wt) and 10 wt% MeOH using 0.5 wt% immobilized lipase, resulting in efficient esterification of FFA. The reaction mixture after 24 h was composed of 49.1 wt% TAG, 1.3 wt% FFA, 49.1 wt% FAME, and negligible amounts of DAG and MAG (<0.5 wt%). The reaction mixture was then dehydrated and used as a substrate for the second reaction, which was conducted at 30°C in a solution of the dehydrated mixture and 5.5 wt% MeOH using 6 wt% immobilized lipase. The activity of the lipase increased gradually when the reaction was repeated by transferring the enzyme to a fresh substrate mixture. The activity reached a maximum after 6 cycles, and the content of FAME achieved was >98.5 wt% after a 24-h reaction. The immobilized lipase was very stable in the first-and second-step reactions and could be used for >100 d without significant loss of activity.     

Enrichment of arachidonic acid: Selective hydrolysis of a single-cell oil fromMortierella withCandida cylindracea lipase

Three lipases, isolated previously in our laboratory, and a known lipase fromCandida cylindracea were screened for the enrichment of arachidonic acid (AA). The enzyme fromC. cylindracea was the most effective for the production of oil with high concentration of AA. When a single-cell oil fromMortierella alpina, containing 25% AA, was hydrolyzed with this lipase for 16 h at 35°C, the resulting glycerides contained 50% AA at 52% hydrolysis. After this, no further hydrolysis occurred, even with additional lipase. However, when the glycerides were extracted from the hydrolyzate and were hydrolyzed again with new lipase, the resulting oil contained 60% AA, with a recovery of 75% of its initial AA content. Triglycerides were the main components of the resulting oil. The release of each fatty acid from the oil depended on the hydrolysis rate of its ester. The fatty acid, whose ester is the poorest substrate for the enzyme, is concentrated in the glycerides.     

Development of a two-dimensional dual pendulum thrust stand for Hall thrusters

A two-dimensional dual pendulum thrust stand was developed to measure thrust vectors [axial and horizontal (transverse) direction thrusts] of a Hall thruster. A thruster with a steering mechanism is mounted on the inner pendulum, and thrust is measured from the displacement between inner and outer pendulums, by which a thermal drift effect is canceled out. Two crossover knife-edges support each pendulum arm: one is set on the other at a right angle. They enable the pendulums to swing in two directions. Thrust calibration using a pulley and weight system showed that the measurement errors were less than 0.25 mN (1.4%) in the main thrust direction and 0.09 mN (1.4%) in its transverse direction. The thrust angle of the thrust vector was measured with the stand using the thruster. Consequently, a vector deviation from the main thrust direction of +/-2.3 degrees was measured with the error of +/-0.2 degrees under the typical operating conditions for the thruster.     

Characteristics of humic substances in the Kuji River waters as determined by high-performance size exclusion chromatography with fluorescence detection

Direct measurement by high-performance size exclusion chromatography with fluorescence detection was applied to the characterization of humic substances in river waters from the Kuji River system, which runs through forest hills and an agricultural plain in Japan. The monitoring wavelength of excitation 320 nm and emission 430 nm corresponds to the fluorescence maxima for aquatic fulvic acid. Chromatograms of the river waters showed four peaks; each peak position was in good agreement among these samples. Peak height ratios for the samples from the upstream Kuji River and its tributaries were different from those of the midstream and downstream sections of the Kuji River, which may reflect differences in the characteristics of humic substances and other organic materials supplied from soil to river.     

Thermal stress measurement in steel plate strengthened by CFRP and aluminum alloy plates

In steel members strengthened by carbon-fiber reinforced polymer (CFRP) plates, the thermal stresses are introduced in the steel members, the CFRP plates and the adhesive layers when temperature changes because the linear thermal expansion coefficients of steel and CFRP are mismatched. As so far, the authors proposed a technique to reduce the thermal stress in steel members strengthened by CFRP plates, which involves bonding aluminum alloy plates with CFRP plates. In the proposed method, the thermal stress in steel member can be reduced so that there are negligible levels of stress in steel member when the cross sectional areas of CFRP and aluminum plates are designed to correspond to the coefficient of thermal expansion of steel, even though the thermal stresses are introduced in the CFRP and aluminum plates. In this study, to confirm the maintaining the thermal stress reduction in steel member by proposed method, thermal stress measurement in steel plate strengthened by CFRP and aluminum plates was carried out about 21 months. In this research, the thermal stress introduced in the steel plate strengthened by CFRP plates was also measured. Furthermore, to assume the thermal shear and normal (peel) stresses in adhesive layers, FE analysis with plane stress element was employed. As the result, it was shown the thermal stresses in steel plate with CFRP plate were able to calculate by using composite theory and measured temperature. Furthermore, in steel plate strengthened by CFRP and aluminum plates, the thermal stress introduced in steel plate was negligible-small through the all-season. It was found the thermal stresses in steel plate with CFRP plates as well as CFRP and aluminum plates were also estimated by using composite theory and measured temperature. In the steel plate strengthened by CFRP and aluminum plates, the thermal shear and normal stresses in adhesive layer glued to steel plate become smaller than that in the conventional CFRP bonded specimen. However, the shear stress in adhesive layers between CFRP and aluminum plates in proposed method was higher than the thermal stress in adhesive layers between CFRP plates in conventional method.     

Nonspecific effects of the pharmacological probes commonly used to analyze signal transduction in rabbit parietal cells

In order to examine some possibly misleading conclusions of the pharmacological analysis of the signal transduction pathways of gastric acid secretion, we evaluated various agents including inhibitors of protein kinase C, cyclic AMP-dependent protein kinase, phospholipase C, phospholipase A2, lipoxygenase, casein kinase, calmodulin, myosin light chain kinase, tyrosine kinase, anion exchanger, and protein phosphatase; and activators of protein kinase C. Among them, the cyclic AMP-dependent protein kinase inhibitor N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinylsulfonamide (H-89), the phospholipase A2 inhibitor 2-(p-amylcinnamoyl)amino-4-chlorobenzoic acid (ONO-RS-082), three myosin light chain kinase inhibitors (1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine (ML-7), 1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine (ML-9), and wortmannin), the anion exchanger inhibitor 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), the phospholipase C inhibitor neomycin, and most known calmodulin antagonists strongly inhibited [14C]aminopyrine accumulation, an indicator of acid secretion, in isolated rabbit gastric glands stimulated by N6,2'-O-dibutyryl-cyclic AMP. ONO-RS-082, calmidazolium, and DIDS inhibited H+,K+-ATPase. Most of the chemicals with antisecretory activity showed protonophore-like activity in gastric microsomes as well as in the mitochondria. It is concluded that H-89, ONO-RS-082, ML-7, ML-9, neomycin, and all calmodulin antagonists tested so far should not be used as tools to analyze gastric acid secretion.     

An integrated approach to the study of living cells by atomic force microscopy

We describe a technique for studying living cells with the atomic force microscope (AFM) in tapping mode using a thermostated, controlled-environment culture system. We also describe the integration of the AFM with bright field, epifluorescence and surface interference microscopy, achieving the highest level of integration for the AFM thus far described. We succeeded in the continuous, long-term imaging of relatively flat but very fragile cytoplasmic regions of COS cells at a lateral resolution of about 70 nm and a vertical resolution of about 3 nm. In addition, we demonstrate the applicability of our technology for continuous force volume imaging of cultured vertebrate cells.
The hybrid instrument we describe can be used to collect simultaneously a diverse variety of physical, chemical and morphological data on living vertebrate cells. The integration of light microscopy with AFM and steady-state culture methods for vertebrate cells represents a new approach for studies in cell biology and physiology.     

Measurements of acoustic waves generated by electrical breakdown in a polypropylene film

Electrical breakdown of a polymer film generates a pressure wave that is believed to include information about the breakdown initiation point. We measured the breakdown pressure wave and the space charge distribution up to the electrical breakdown field by using the pulsed electro‐acoustic method in a 30 µm thick polypropylene film. We discuss electrical breakdown phenomena based on the breakdown pressure wave and the dependence of the space charge distribution on the applied field and temperature. At room temperature, the observed breakdown pressure wave had a pulse‐like shape with a width that depended on the polarity of the applied field. Positive space charge accumulation was observed near the cathode as a hetero space charge near the electrical breakdown field. At 60 °C, the width of the breakdown pressure wave showed no dependence on the applied field polarity and positive space charge accumulation was observed inside the film near the electrical breakdown field. These experimental results suggest that electrical breakdown phenomena are affected by hetero space charge accumulation and that the initiation point of electrical breakdown corresponds to the position of hetero space charge accumulation in 30 µm thick polypropylene film. ©1999 Scripta Technica, Electr Eng Jpn, 126(3): 1–8, 1999     

Fabrication of barium titanate nanoparticles‐epoxy resin composite films and their dielectric properties

A method for fabricating epoxy resin films dispersing the surface‐modified barium titanate (BT) particles (BT‐epoxy resin composite films) are proposed. BT particles with a size of 7.8 nm and a crystal size of 8.6 nm were synthesized with a complex alkoxide method. To introduce epoxy groups on the BT particle surface, the BT particles were surface‐modified with 2‐(3,4‐epoxycyclohexyl)‐ethyltrimethoxysilane. A precursor solution, which was prepared by prereacting 2,2‐bis(4‐glycidyloxyphenyl)propane (BGPP) and phthalic anhydride in 4‐butyrolactone and adding the surface‐modified BT particles to the prereacting solution, was spin‐coated on glass substrates to fabricate the composite films. An increase in BT volume fraction in film increased dielectric constant of the composite film while keeping dissipation factor below 0.03. The dielectric constant attained 10.8 at a BT volume fraction of 30% in film that was around twice higher than pure epoxy resin film. POLYM. COMPOS., 31:1179–1183, 2010. © 2009 Society of Plastics Engineers     

Coarse-grained molecular dynamics simulations on size effect of glassy polyethylene particles

The size effect on mechanical properties of glassy polyethylene (PE) nanoscale particles has been investigated by extensive coarse-grained molecular dynamics simulations. The diameter of the PE particles varies in the range of 5-40 nm, we confirm that the particle's behaviour under compressive stress strongly depends on its size-the smaller the particle diameter is, the stiffer the particle behaves. The present mechanical responses of compressed particles are in good agreement with our previous experimental phenomena of micron-sized polymer particles measured by a nanoindentation-based flat punch method. Possible reasons for the size effect are discussed.