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.     

Modification of the analysis of parathyroid hormone-related protein in milk and concentrations of this protein in commercial milk and milk products in Japan

Parathyroid hormone-related protein (PTHrP), which causes hypercalcemia associated with malignant tumors, is known to be present in milk. Gene expression of PTHrP in the mammary gland increases markedly during parturition and with the onset of lactation. Even when circulating PTHrP levels are extremely low or below the detection limit, milk PTHrP levels are remarkably high. Parathyroid hormone-related protein derived from the mammary gland is assumed to play a role in maintaining the maternal calcium homeostasis and calcium transport from blood to milk. In previous studies that determined the PTHrP concentrations in milk, the pretreatments and diluent composition were not standardized. Here, we investigated the effect of various pretreatment procedures and diluent constitutions and the consequent PTHrP concentrations in commercial milk and milk products in Japan. Significant differences were found in PTHrP concentrations in raw milk samples subjected to different combinations of pretreatments (mixing, centrifugation, acidification, and heating) and diluents (0 pM standard solution of PTHrP, plasma treated with protease inhibitors, and original diluent). We measured the PTHrP concentrations in normal liquid milk, processed milk, milk drinks, formulated milk powders, and skim milk powder by using the appropriate combination of pretreatment (acidification) and diluent (plasma treated with protease inhibitors). The PTHrP concentration in normal liquid milk, processed milk, and skim milk powder was as high as that in raw milk (>5 nM), whereas that in milk drinks differed considerably. The PTHrP concentration in infant formulas (<2 nM) was lower than that in the other milk products. These results indicate that a certain amount of PTHrP is ingested when milk and milk products are consumed.     

Role of peritoneal lavage smears and gastric wall brushing smears in gastric cancer surgery

Examinations of peritoneal lavage smears (LC) and gastric wall brushing smear cytology (BC) appear to be important for determining accurately the stage of gastric cancer. We have been carrying out such examinations during gastric cancer surgery for 7 years. In the present study, we evaluated the results obtained from 287 patients with gastric cancer. Tumor invasion and peritoneal dissemination were correlated with a positive incidence of cancer cells in LC and/or BC. Gastric cancer showing serosal invasion was classified into positive for LC and/or BC and negative for LC and/or BC. Patients positive for LC and/or BC had a poorer prognosis. For future gastric cancer treatment, patients positive for peritoneal cytology are expected to be targeted for intensive treatment during or before surgery.     

Extraction of a plasma time-activity curve from dynamic brain PET images based on independent component analysis

A compartment model has been used for kinetic analysis of dynamic positron emission tomography (PET) data [e.g., 2-deoxy-2-18F-fluoro-D-glucose (FDG)]. The input function of the model [the plasma time-activity curve (pTAC)] was obtained by serial arterial blood sampling. It is of clinical interest to develop a method for PET studies that estimates the pTAC without needing serial arterial blood sampling. For this purpose, we propose a new method to extract the pTAC from the dynamic brain PET images using a modified independent component analysis [extraction of the pTAC using independent component analysis (EPICA). Source codes of EPICA are freely available at http://www5f.biglobe.ne.jp/?kimura/Software/top.html]. EPICA performs the appropriate preprocessing and independent component analysis (ICA) using an objective function that takes the various properties of the pTAC into account. After validation of EPICA by computer simulation, EPICA was applied to human brain FDG-PET studies. The results imply that the EPICA-estimated pTAC was similar to the actual measured pTAC, and that the estimated blood volume image was highly correlated with the blood volume image measured using 15O-CO inhalation. These results demonstrated that EPICA is useful for extracting the pTAC from dynamic PET images without the necessity of serial arterial blood sampling.     

Hydrolysis of Cellulose Selectively into Glucose Over Sulfonated Activated-Carbon Catalyst Under Hydrothermal Conditions

A sulfonated activated-carbon (AC-SO₃H) catalyst selectively hydrolyzes cellulose with β-1,4-glycosidic bonds into glucose in the catalytic hydrothermal reactions at temperatures around 423 K. The AC-SO₃H catalyst with the hydrothermal pre-treatment has the excellent catalytic properties attributed to the high hydrothermal stability and the strong acid sites of sulfo functional groups.     

A Restart Control Method for Position Sensorless PMSM Drive Systems Without Potential Transformer for Railway Vehicle Traction

This paper proposes a restart control method for position sensorless PMSM drive systems without a potential transformer for railway vehicle traction. This method can estimate the initial rotor speed and position under coasting conditions over the entire speed range. The method can also be used when the back‐EMF voltage is higher than the inverter DC link voltage. The proposed method is verified by experiments using a 200‐kW PMSM.     

High-Intensity Discharge Lamp with Mo–SiO2 Functionally Graded Material

We show the invention of the new type of hermetically sealed high-intensity discharge lamps, made of Mo–SiO₂ functionally graded material (FGM) as an electrode and a sealing component. In the case of high-intensity discharge lamps with Mo–SiO₂ FGM (FGM lamp), the thermal expansion coefficient between Mo and SiO₂ is functionally graded so that it tolerates a large number of heating cycles, with no cooling system required. Furthermore, lamps survive without breakage. The W electrode is totally separated from the lamp envelope by the FGM, so that no leakage of the luminous elements or gases takes place, when a large gas pressure exists inside a lamp.     

Photo-thermally cured eugenol-derived epoxy resins by simultaneous thiol-ene/thiol-epoxy/thiol-maleimide triple “click” reactions

Polyglycidyl ether of eugenol novolac (PGEEGN) was synthesized by the glycidylation reaction of eugenol novolac (EGN) with an average degree of polymerization of ca. 3. A mixture of PGEEGN and a pentaerythritol-based tetrathiol (S4P) was photo-polymerized at room temperature and subsequently thermally cured at 100–150 °C to produce a two-component cured product (PGEEGN-S4P). A similar curing reaction of glycidyl ether of eugenol (GEEG) and S4P produced another two-component cured product (GEEG-S4P). Furthermore, a mixture of PGEEGN, S4P and 4,4′-bismaleimidodiphenylmethane (BMI) was photo-polymerized at room temperature and subsequently thermally cured at 100–230 °C to produce a three-component cured product (PGEEGN-S4P-BMI). The FT-IR spectral analysis revealed that the thiol-ene and thiol-epoxy reactions progressed for GEEG-S4P and PGEEGN-S4P, and the thiol-ene, thiol-epoxy and thiol-maleimide reactions progressed for PGEEGN-S4P-BMI. The 5% weight loss temperatures of PGEEGN-S4P and PGEEGN-S4P-BMI were higher than that of GEEG-S4P. A higher order of T_g, tensile strength and modulus was PGEEGN-S4P-BMI?>?PGEEGN-S4P?>?GEEG-S4P. The oligomerization of eugenol units and incorporation of BMI were effective to improve thermal and mechanical properties of the GEEG/S4P curing system.