E-cadherin gene-engineered feeder systems for supporting undifferentiated growth of mouse embryonic stem cells

Conventionally, embryonic stem (ES) cells are cultured on a cell layer of mouse embryonic fibroblasts (MEFs) as feeder cells to support undifferentiated growth of ES cells. In this study, cell–cell interactions between mouse ES and feeder cells were artificially engineered via an epithelial cell adhesion molecule, E-cadherin, whose expression is considerable in ES cells. Mouse mesenchymal STO and NIH3T3 cells that were genetically engineered to express E-cadherin were used in ES cell cultures as feeder cells. ES cells cultured on the E-cadherin-expressing feeder cells maintained the expression of stem cell markers, alkaline phosphatase (AP), Oct3/4, Nanog and Sox2, and the efficiency of AP-positive colony formation was comparable to MEFs, and much better than parental STO and NIH3T3 cells. Furthermore, ES cells maintained on the E-cadherin-expressing feeder cells possessed the ability to differentiate into the three germ layers both in vitro and in vivo. The results indicated that E-cadherin expression in feeder cells could improve the performance of feeder cells, which may be further applicable to create new artificial feeder cell lines.     

Modeling of diffusive mass transport in micropores in cement based materials

In order to predict long-term leaching behavior of cement constituents for safety assessments of radioactive waste disposal, we modeled diffusive mass transport in micropores in cement based materials. Based on available knowledge on the pore structure, we developed a transport porosity model that enables us to estimate effective porosity available for diffusion (transport porosity) in cement based materials. We microscopically examined the pore structure of hardened cement pastes to partially verify the model. Effective diffusivities of tritiated water in hardened cement pastes were also obtained experimentally, and were shown to be proportional to the estimated transport porosity.     

Validation of MAAP model enhancement for Fukushima Dai-ichi accident analysis with Phenomena Identification and Ranking Table (PIRT)

The Modular Accident Analysis Program (MAAP) model enhancement items to improve the simulation capability for molten corium behavior in the accidents at the Fukushima Dai-ichi nuclear power plants were validated with the Phenomena Identification and Ranking Table (PIRT). The importance ranks of the identified phenomena were evaluated for each time phase through brainstorming and discussion with the experts in the Atomic Energy Society of Japan and the members of the MAAP model enhancement project. When the current MAAP evaluation models were reviewed with the PIRT, it is found that 95 of the 386 high-ranked phenomena were not considered in MAAP 5.0.1. While 62 of these phenomena will have been addressed in the MAAP enhancement project and 25 others are not suitable to be analyzed by MAAP, 8 important phenomena should be considered in post-MAAP enhancement project with additional experiments or fundamental studies.     

Sequential exposure of porcine cumulus cells to FSH and/or LH is critical for appropriate expression of steroidogenic and ovulation-related genes that impact oocyte maturation in vivo and in vitro

In this study, we collected follicular fluid, granulosa cells, and cumulus cells from antral follicles at specific time intervals following equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) treatment of gilts. The treatment with eCG increased the production of estrogen coordinately with up-regulated proliferation of granulosa and cumulus cells. eCG also induced the expression of LHCGR and PGR in cumulus cells and progesterone accumulation was detected in follicular fluid prior to the LH/hCG surge. Moreover, progesterone and progesterone receptor (PGR) were critical for FSH-induced LHCGR expression in cumulus cells in culture. The expression of LHCGR mRNA in cumulus cells was associated with the ability of LH to induce prostaglandin production, release of epidermal growth factor (EGF)-like factors, and a disintegrin and metalloprotease with thrombospondin-like repeats 1 expression, promoting cumulus cell oocyte complexes (COCs) expansion and oocyte maturation. Based on the unique expression and regulation of PGR and LHCGR in cumulus cells, we designed a novel porcine COCs culture system in which hormones were added sequentially to mimic changes observed in vivo. Specifically, COCs from small antral follicles were pre-cultured with FSH and estradiol for 10 h at which time progesterone was added for another 10 h. After 20 h, COCs were moved to fresh medium containing LH, EGF, and progesterone. The oocytes matured in this revised COC culture system exhibited greater developmental competence to blastocyst stage. From these results, we conclude that to achieve optimal COC expansion and oocyte maturation in culture the unique gene expression patterns in cumulus cells of each species need to be characterized and used to increase the effectiveness of hormone stimulation.     

Effect of addition of corrosion inhibitors on corrosion behaviors of PCV material under diluted artificial seawater environment

ABSTRACT

Fukushima Daiichi nuclear power plants (1F) were damaged by unprecedented severe accident in the great east Japan earthquake, and seawater and freshwater had been injected as an emergency countermeasure for the core cooling. Although, the primary containment vessel (PCV) was not supposed to be exposed to diluted seawater, the PCV will be exposed to diluted seawater environment until fuel debris removal is completed. Therefore, it is necessary to establish a countermeasure of corrosion for the PCVs made of carbon steel. In this study, the effect of the addition of corrosion inhibitor, which is one of the corrosion countermeasures, was investigated by two types of corrosion tests. As a result of the immersion corrosion test, it was found that any of the three kinds of corrosion inhibitor could suppress corrosion of carbon steel. In addition, as a result of the inhibitor interim addition test, it was found that corrosion of carbon steel covered with corrosion products could be suppressed by optimizing the additive amount of corrosion inhibitor in the cooling water.     

Long-term effects of a selective alpha 1-adrenergic inhibitor on right and left ventricular masses in patients with chronic pulmonary disease]

The effects of pinacidil and nimodipine on endothelin-1-induced contractions in isolated cerebral arteries with and without endothelium were compared. The sensitivity to endothelin-1 was increased (0.5 log units) in the rabbit basilar artery after removal of the endothelium. The nitric oxide synthase inhibitor N omega-nitro-L-arginine (0.1 mM) also increased the sensitivity to endothelin-1 (0.6 log units) in basilar arteries with endothelium, whereas N omega-nitro-D-arginine (0.1 mM) and indomethacin (3 microM) had no effect, indicating that withdrawal of endothelium-derived nitric oxide may account for the enhancement of the endothelin-1-induced contraction after endothelial denudation. Pinacidil (1 microM) shifted the concentration-response curve for endothelin-1 to the right without affecting the maximal response in arteries without endothelium, but had no effect on the endothelin-1-induced contraction in vessels with endothelium. Nimodipine (1 microM) reduced the maximal endothelin-1-induced contraction by approximately 50% in both the presence and absence of endothelium, whereas the sensitivity to endothelin-1 was reduced only in vessels without endothelium. Incubation in "calcium-free" medium reduced the maximal endothelin-1-induced contraction by 69% and 80% in vessels with and without endothelium, respectively. In human pial arteries with endothelium, pinacidil did not affect the endothelin-1-induced contraction, whereas nimodipine and exposure to "calcium-free" solution reduced the maximal response by 31% and 74% respectively. The results show that, in the rabbit, pinacidil and to a lesser extent nimodipine preferentially act on cerebral arteries with disrupted endothelium, indicating that vasoactive factors liberated from the endothelium may modify the effect of a vasodilator.     

Actions of taurine on the L-type Ca2+ channel current in guinea pig ventricular cardiomyocytes

This article presents a new method for analyzing the spreading of skin erythemas. These occur as a result of the cutaneous vascular axon reflex which can be evoked by a noxious stimulation of the skin. Series of true-color images of the observed skin patch were recorded using a video camera. The images were digitized and stored on computer disk. The delineation of the reddening was segmented for every image of the sequence by a newly developed image processing method. Each image taken after the noxious stimulation was compared with the baseline before the stimulation and each image point was classified as: "unchanged" or "changed skin color." To improve the classification the CIE L*a*b* color space was used. The boundaries of the erythema were extracted from the resulting binary images. Every image of a sequence was analyzed in the same way in order to follow the time course of the flare response. The erythema reaction could be determined in an objective way using this methods. The automatically detected flare sizes were independent of human observers and had a high spatial and temporal resolution. It was used for a crossover study to assess the power of new drugs which modify the blood flow of the skin induced by an intradermal histamine application.     

Estimating leaf carotenoid contents of shade-grown tea using hyperspectral indices and PROSPECT–D inversion

Quantifying carotenoid contents has many applications in agriculture, ecology, and health science. Hyperspectral reflectance has been one of the promising tools for this purpose. However, previous studies were based on measurements under relatively low light–stress conditions. Therefore, assessing its robustness by using measurements under various levels of stress is required. In this study, the measurements of reflectance and carotenoid contents were carried out with four shading treatments including open–0%, 35%, 75%, and 90% shading to generate various chlorophyll/carotenoid ratios. Then the performances of 15 published hyperspectral indices and PROSPECT–D inversion were evaluated based on our data set for estimating leaf carotenoid contents. According to the ratio of performance to deviation, R_NIR/R₅₁₀, R₇₂₀/R₅₂₁–1, and PROSPECT–D inversion were applicable for this purpose, although calibration of the absorption coefficients was required for PROSPECT–D. Using them, root mean square percentage errors of 4.53–5.46% were achieved. Given that total chlorophyll/carotenoid ratios could be a good indicator for evaluating environmental stress in plants, PROSPECT–D, which also estimates total chlorophyll and anthocyanin contents, could be a strong tool for controlling the qualities of shade-grown tea.     

Consolidation of Sm5Fe17 powder by spark plasma sintering method

Sm₅Fe₁₇ bulk materials, one of the newer permanent magnetic materials, were successfully produced by the spark plasma sintering method. The resultant bulk materials had high densities of 85–98%. When obtained by sintering at relatively lower temperatures, the Sm₅Fe₁₇ bulk materials consisted of the Sm₅Fe₁₇ and SmFe₃ phases, whereas they contained some -Fe phase together with the Sm₅Fe₁₇ and SmFe₃ phases when obtained by sintering at relatively high temperatures. High coercivity values, exceeding 2 MAm⁻¹, were found in the Sm₅Fe₁₇ bulk materials consisting of the Sm₅Fe₁₇ and SmFe₃ phases.