Contribution of high p34cdc2 kinase activity to premature chromosome condensation of injected somatic cell nuclei in rat oocytes

The present study was undertaken to clarify the relationship between the p34cdc2 kinase activity of in vitro-aged or enucleated rat oocytes and the premature chromosome condensation (PCC) of microinjected cumulus cell nuclei. Wistar rat oocytes were placed in vitro up to 120 min after the animal was killed. The p34cdc2 kinase activity of the oocytes decreased in a time-dependent manner. The incidence of PCC was higher when nuclear injection into intact oocytes was completed in 15-45 min rather than 46-120 min. When rat oocytes were enucleated for subsequent nuclear injection, the p34cdc2 kinase activity transiently increased soon after enucleation but drastically decreased after 30 min. Removal of the cytoplasm instead of the meta-phase-plate did not affect the p34cdc2 kinase activity even after 60 min. PCC occurred in intact and cytoplasm-removed oocytes but not in enucleated oocytes. In contrast, oocytes from BDF1 mice exhibited a p34cdc2 kinase level twice that of rat oocytes and supported PCC despite enucleation. The p34cdc2 kinase level of intact rat oocytes was reduced to the equivalent level of aged (120 min) or enucleated (+60 min) oocytes by a 45 min treatment with roscovitine, an inhibitor of p34cdc2 kinase. None of the roscovitine-treated oocytes supported PCC while half of the control oocytes did. When rat oocytes were treated with MG132, a proteasome inhibitor, delayed inactivation of the p34cdc2 kinase was observed in the MG132-treated oocytes. A significantly higher proportion of the MG132-treated oocytes supported PCC when compared with the control oocytes. Moreover, a higher proportion of MG132-treated and enucleated oocytes carried two pseudo-pronuclei after cumulus cell injection and developed to the two-cell stage when compared with the enucleated oocytes at the telophase-II stage. These results suggest that the decreased level of p34cdc2 kinase activity in aged or enucleated rat oocytes is responsible for their inability to support PCC of microinjected donor cell nuclei and that inhibition of p34cdc2 kinase inactivation by chemicals such as MG132 is in part effective for rat oocytes to promote PCC and further development.     

Carbonization and liquid—crystal (mesophase) development. 19. Co-carbonization of oxidized coals with model organic compounds

This study examines further the phenomena of the modification of coal carbonizations by organic additives. Anthracene, pyrene and chrysene modify the carbonization in a closed system of coking coals as observed from increases in the size of optical textures of resultant cokes. Weakly caking coals are unaffected. Chrysene is the most efficient modifier probably because of its lowest calculated free valence. The co-additives tetralin and hydrogenated anthracene oil further enhance the modification processes so obviating the necessity to use hydrogenated additives. Co-carbonizations of oxidized coking and caking coals with decacyclene are effective in removing the effects of mild oxidation. Increased rates of carbonization enhance the sizes of optical textures of resultant cokes.     

Determining surface orientations of transparent objects based on polarization degrees in visible and infrared wavelengths

Techniques for modeling an object through observation are very important in object recognition and virtual reality. A wide variety of techniques have been developed for modeling objects with opaque surfaces, whereas less attention has been paid to objects with transparent surfaces. A transparent surface has only surface reflection; it has little body reflection. We present a new method for obtaining surface orientations of transparent surfaces through analysis of the degree of polarization in surface reflection and emission in visible and far-infrared wavelengths, respectively. This parameter, the polarization degree of reflected light at the visible wavelengths, is used for determining the surface orientation at a surface point. The polarization degree at visible wavelengths provides two possible solutions, and the proposed method uses the polarization degree at far-infrared wavelengths to resolve this ambiguity.     

Effect of aging on microstructural development in an Al–Mg–Si alloy processed by high-pressure torsion

Experiments were conducted to evaluate the microstructural evolution in a commercial Al-0.6 % Mg-0.4 % Si alloy processed using high-pressure torsion for up to 20 turns. Disks of the alloy were tested in two different conditions: in a solution-treated condition and after a short aging treatment at 523 K. The results show that HPT processing introduces significant grain refinement through HPT processing including a reduction in grain size from ~150 μm to ~720 nm in 1 turn of HPT. The final grain size in this alloy was ~250 nm after 20 turns. Some tensile tests were conducted to evaluate the mechanical properties of the alloy at the solution treatment temperature. The results from these tests show that aging at 523 K leads to a small increase in ductility for all tensile samples with a maximum recorded elongation of ~230 %.     

Different models of hardness evolution in ultrafine-grained materials processed by high-pressure torsion

High-pressure torsion (HPT) is an attractive processing method in severe plastic deformation techniques involving the application of high compressive pressure with concurrent torsional straining. Excellent grain refinement is anticipated when using this technique to average grain sizes of the submicrometer or even nanometer ranges. Because of the significant microstructural changes during processing, there are numerous reports showing evolution in local hardness toward homogeneity throughout a disk diameter with increasing numbers of revolutions. The achieved hardness after HPT is mostly much higher than that in the as-received condition because of exceptional grain refinement although there are a limited number of metals and alloys showing softening or weakening after HPT processing. This paper was initiated to review recent discoveries in the experimental results on hardness evolution toward homogeneity during HPT processing and discuss the different models of hardness developments with respect to imposed equivalent strain by HPT processing for a range of metals and alloys. Moreover, recent results of hardness homogeneity and heterogeneity through thicknesses of the processed disks are discussed toward a complete understanding of hardness evolution in the UFG metals processed by HPT.     

Applicability of DNA barcode for identification of fish species

DNA barcoding is a species identification technique, which uses a very short DNA sequence from a region of approximately 650 base-pairs in the 5'-end of the mitochondrial cytochrome c oxidase subunit I gene as a marker to identify species of mammals and fishes. The applicability of DNA barcoding for identification of fish species consumed in Japan was studied. Among thirty-one fresh or processed fishes were obtained from the market, two samples could not be identified due to lack of data in the Barcode of Life Data (BOLD) database. However, BLAST-search of 16S rRNA genes in the National Center for Biotechnology Information (NCBI) database and the PCR-RFLP method published by the Food and Agricultural Materials Inspection Center (FAMIC) were found to be applicable to identify these 2 fishes. The results show that the DNA barcoding technique is potentially useful as a tool for confirming the proper labeling of fish species in the Japanese market.     

Stability of the ultrafine-grained microstructure in silver processed by ECAP and HPT

The high-temperature thermal stability of the ultrafine-grained (UFG) microstructures in low stacking fault energy silver was studied by differential scanning calorimetry (DSC). The UFG microstructures were achieved by equal-channel angular pressing (ECAP) and high-pressure torsion (HPT) at room temperature (RT). The defect structure in the as-processed samples was examined by electron microscopy and X-ray line profile analysis. The stored energy calculated from the defect densities was compared to the heat released during DSC. The sum of the energies stored in grain boundaries and dislocations in the ECAP-processed samples agreed with the heat released experimentally within the experimental error. The temperature of the DSC peak maximum decreased while the released heat increased with increasing numbers of ECAP passes. The released heat for the specimen processed by one revolution of HPT was much smaller than after 4–8 passes of ECAP despite the 2 times larger dislocation density measured by X-ray line profile analysis. This dichotomy was caused by the heterogeneous sandwich-like microstructure of the HPT-processed disk: about 175 μm wide surface layers on both sides of the disk exhibited a UFG microstructure while the internal part was recrystallized, thereby yielding a relatively small released heat.     

Development of effective detection method for Coxiella burnetii in mayonnaise by real-time PCR and investigation of C. burnetii contamination in commercial mayonnaise in Tokyo

A PCR method for the effective detection of Coxiella burnetii in commercially available mayonnaise was developed. Sample preparations were isolated from 50 g portions of each mayonnaise product by four successive extraction steps in phosphate buffer with 2.0 M NaCl. These extracts were then centrifuged at 20,000 x g for 60 min. DNA was isolated from the solution containing the precipitate with a commercial kit, and amplified quantitatively using real-time PCR that targeted the com1 region of C. burnetii. The recoveries of C. burnetii from 2 kinds of commercial mayonnaise specimens, with a baseline control of 1 x 10(7) particles of the Nine Mile phase II strain, were 85.0 +/- 6.0% and 72.0 +/- 0.4%, respectively. The determination limit of this method was 500 C. burnetii particles per 50 g of mayonnaise. The DNA specimens isolated from 50 different commercial mayonnaise samples sold in Tokyo using this method were amplified using both nested PCR and real-time PCR. No contamination by C. burnetii was detected in any of the mayonnaise samples.