Liberation of phospholipids from Z-disks of chicken skeletal muscle myofibrils by 0.1 mM calcium ions: weakening mechanism for Z-disks during post-mortem aging of meat

Weakening of the Z-disks of skeletal muscle myofibrils contributes to the tenderization of meat during post-mortem aging. To elucidate the weakening mechanism, we compared Z-disks weakened by post-mortem aging of chicken breast muscle with those of myofibrils treated with a solution containing 0.1 mM CaCl2 and 1 microM calpastatin domain I. In both cases, the Z-disks were weakened with a corresponding liberation of their constituent phospholipids (PLs). The liberation of PLs specific to 0.1 mM calcium ions was minimal at pH 6.5 and maximal at 35 degrees C together with the Z-disk weakening. Binding of calcium ions to PLs in the Z-disks was determined by 45Ca-autoradiography. Acidic PLs were strongly radioactive and neutral PLs were appreciably radioactive. It is very probable that acidic PLs would bind electrostatically to alpha-actinin under physiological conditions, and that this interaction would be broken by the binding of calcium ions at 0.1 mM to PLs, resulting in the partial liberation of PLs from Z-disks. We conclude, therefore, that the liberation of PLs by the binding of 0.1 mM calcium ions was the main cause for Z-disk weakening during the post-mortem aging of chicken.     

A cis-regulatory element within the 5'' flanking region of arylsulfatase gene of sea urchin, Hemicentrotus pulcherrimus

Mycobacteria generally have thick cell walls and contain large amounts of lipid, making them resistant to DNA extraction. Five methods, namely, extensive enzymic digestion method (M1), 2-min mechanical glass-bead disruption method (M2), thermal shock method (M3), modified conventional enzymic digestion method (M4), and manual disruption with modified conventional enzymic digestion method (M5), were used to compare their effectiveness and simplicity in extracting DNA from slowly growing mycobacteria (Mycobacterium leprae, M. lepraemurium and M. bovis BCG), and a rapidly growing mycobacterium (M. phlei). The highest DNA yield was obtained by M2 from M. lepraemurium which produced 2.82 micrograms DNA/mg wet weight of cells, representing a theoretical yield of 78%. M3 gave the lowest DNA yield; 0.01 microgram DNA/mg wet weight of cells of M. lepraemurium was obtained. M4, in which proteinase K was used, is more effective than M1, in which subtilisin and pronase were used. M5 yielded a higher amount of DNA, but it required more manipulations to extract DNA as compared to M4. Extraction of DNA of M. leprae from nude mice is more difficult than that of M. leprae from armadillos by all of the methods used. These results suggest that the biosynthetic capabilities of these two forms of M. leprae may vary, depending on their cultural conditions and/or strain differences. Our results have shown that both M2 and M4 are the simplest, most effective and time-saving methods which are suitable for every routine laboratory to extract DNA from slowly and rapidly growing mycobacteria.     

Preparation of regioisomers of structured TAG consisting of one mole of CLA and two moles of caprylic acid

TAG (MLM) with medium-chain FA (MCFA) at the 1,3-positions and long-chain FA (LCFA) at the 2-position, and TAG (LMM) with LCFA at the 1(3)-position and MCFA at 2,3(1)-positions are a pair of TAG regioisomers. Large-scale preparation of the two TAG regioisomers was attempted. A commercially available FFA mixture (FFA-CLA) containing 9-cis, 11-trans (9c, 11t)- and 10t,12c-CLA was selected as LCFA, and caprylic acid (C₈FA) was selected as MCFA. The MLM isomer was synthesized by acidolysis of acyglycerols (AG) containing two CLA isomers with C₈FA: A mixture of AG-CLA/C₈ FA (1∶10, mol/mol) and 4 wt% immobilized Rhizomucor miehei lipase was agitated at 30°C for 72 h. The ratio of MLM to total AG was 51.1 wt%. Meanwhile, LMM isomer was synthesized by acidolysis of tricaprylin with FFA-CLA: A mixture of tricaprylin/FFA-CLA (1∶2, mol/mol) and 4 wt% immobilized R. miehei lipase was agitated at 30°C for 24 h. The ratio of LMM to total AG was 51.8 wt%. MLM and LMM were purified from 1,968 and 813 g reaction mixtures by stepwise short-path distillation, respectively. Consequently, MLM was purified to 92.3% with 49.1% recovery, and LMM was purified to 93.2% with 52.3% recovery. Regiospecific analyses of MLM and LMM indicated that the 2-positions of MLM and LMM were 95.1 mol% LCFA and 98.3 mol% C₈ FA, respectively. The results showed that a process comprising lipase reaction and short-path distillation is effective for large-scale preparation of high-purity regiospecific TAG isomers.     

Oxidation kinetics for <Emphasis Type="Italic">cis</Emphasis>-9,<Emphasis Type="Italic">trans</Emphasis>-11 and <Emphasis Type="Italic">trans</Emphasis>-10,<Emphasis Type="Italic">cis</Emphasis>-12 isomers of CLA

The autoxidation processes of the cis-9,trans-11 (c9,t11) and trans-10,cis-12 (t10,c12) isomers of CLA were separately observed at ca. 0% RH and different temperatures. The t10,c12 CLA oxidized faster than the c9,t11 isomer at all tested temperatures. The first half of the oxidation process of t10,c12 CLA obeyed an autocatalytic-type rate expression, but the latter half followed first-order kinetics. On the other hand, the entire oxidation process of c9,t11 CLA could be expressed by the autocatalytic-type rate expression. The apparent activation energies and frequency factors for the autoxidation of the isomers were estimated from the rate constants obtained at various temperatures based on the Arrhenius equation. The apparent activation energies for the CLA isomers were greater than those for the nonconjugated n−6 and n−3 PUFA or their esters. However, the enthalpyentropy compensation held during the autoxidation of both the CLA and PUFA. This suggested that the autoxidation mechanisms for the CLA and PUFA were essentially the same.     

Incorporation of Dust Particles into a Growing Film During Silicon Dioxide Deposition from a TEOS/O2 Plasma

The effects of gas flow rate on particle formation and film deposition during the preparation of silica thin film using a TEOS/O₂ plasma were investigated. Particle formation and growth are suppressed with increasing gas flow rates. The film deposition rate increases with increasing gas flow rate, reaches a maximum value, and eventually decreases again. However, the uniformity of the film tends to degrade at high gas flow rates. At a high gas flow rate, some particles trapped in the sheath near the grounded electrode pass through the sheath to reach the substrate and are then embedded in the growing film. A self-consistent sheath model combined with particle force balance based on charge fluctuation was developed to explain these experimental findings qualitatively. The model reveals that charge fluctuation is a key factor for the particle to overcome the potential barrier of the negatively charged particles to pass through the sheath, eventually reaching the substrate. The model further shows that the probability of a particle being deposited on the substrate is higher for increased gas flow rates, which correctly predicts the experimentally observed trend.     

Unipolar Charging of Nanosized Aerosol Particles Using Soft X-ray Photoionization

A unipolar charging device based on a soft X-ray (<9.5 keV) photoionization was developed to investigate the charging efficiency of aerosol nanoparticles. Unipolar charging using a 241 Am charger was also evaluated as a comparison with the characteristics obtained by X-ray charging. The production rate and the concentration of ions generated by the X-ray and 241 Am unipolar chargers were estimated from ion current measurements. Theoretical calculations by the unipolar diffusion charging theory were also carried out and the calculated data were compared with the experimental results. For acquiring a high number of standard nanoparticles, the classification of monodisperse nanoparticles from polydisperse aerosol particles using the X-ray unipolar charger and a differential mobility analyzer (DMA) was also evaluated. The ion production rate of the X-ray unipolar charger was at least 5.5 times higher than that of the 241 Am unipolar charger and the ion concentration was about three times higher. Therefore, the X-ray unipolar charger showed a higher capability for charging aerosol particles of 10-40 nm size in diameter than the 241 Am charger. The charging state of particles produced by the X-ray unipolar charger was in good agreement with theoretical calculations. The X-ray unipolar charger developed herein has potential for use in charging a high number concentration of nanoparticles for use in nanotechnology investigations.     

Investigation of Ozone Concentration Measurement by Visible Photo Absorption Method

The measurement of ozone concentration based on the visible photo absorption method is investigated. An LED having an emission peak at 609 nm and a photo diode sensitive at visible light frequencies are used as the light source and the photo detector, respectively. The transmittance is measured as a function of ozone concentration, which is in reasonable agreement with the Beer–Lambert relationship. The effective absorption cross-section can therefore be estimated from the fitting procedure. The concentrations determined by both the UV and visible photo absorptions agree well with each other in the range from 7.1 to 68.4 g/m³(N). The relative error in these concentrations is discussed on the basis of the Twyman–Lothian curve. The ozone measurement for the air-fed ozone generation is also investigated. The effect of photo absorption due to NOx species is unavoidable. The present results provide useful data if the ozone measurement based on the visible photo absorption method is applied to ozone monitoring instruments or used for laboratory experiments.     

Novel Fabrication of High-Quality ZrO2 Ceramic Thin Films from Aqueous Solution

We report on the direct deposition of high-quality ZrO₂ thin films on various kinds of substrates by the liquid phase deposition method. After reaction for 24 h, thin films formed on various kinds of substrates, and the obtained thin film was comprised of densely packed nano-sized particles. The film annealed at 500°C showed a tetragonal phase at room temperature and this phenomenon has been discussed from the viewpoint of crystallite size effect. The result of optical transmittance measurement revealed that high transparency, more than 70% transmittance, has been achieved for the film after annealing at 900°C.     

Effect of Nd2O3 Concentration on the Defect Structure of CeO2–Nd2O3 Solid Solution

An extensive X-ray study of CeO₂–Nd₂O₃ solid solutions was performed, and the densities of solid solutions containing various concentrations of NdO_1.5 were measured using several techniques. Solid solutions containing 0–80 mol% NdO_1.5 were synthesized by coprecipitation from Ce(NO₃)₃ and Nd(NO₃)₃ aqueous solutions, and the coprecipitated samples were sintered at 1400°C. A fluorite structure was observed for CeO₂–NdO_1.5 solid solutions with 0–40 mol% NdO_1.5, which changed to a rare earth C-type structure at 45–75 mol% NdO_1.5. The change in the lattice parameters of CeO₂–NdO_1.5 solid solutions, when plotted with respect to the NdO_1.5 concentration, showed that the lattice parameters followed Vegard's law in both the fluorite and rare earth C-type regions. The maximum solubility limit for NdO_1.5 in CeO₂ solid solution was approximately 75 mol%. The relationship between the density and the Nd concentration indicated that the defect structure followed the anion vacancy model over the entire range (0–70 mol% NdO_1.5) of solid solution.