Effects of hormone sources on developmental competence of oocytes by ovum pickup in Japanese black cattle.

Japanese Black (Wagyu) cattle donors were primed with different protocols and sources of follicle-stimulating hormone (FSH) for successive ovum pickup (OPU) and embryo development after in vitro fertilization (IVF). Following OPU, retrieved cumulus oocyte complexes (COCs) were subjected to IVF, and resulting blastocysts were transferred into recipients to evaluate implantation capability. Experiment 1: The best blastocyst development (45.3 %) and embryo yields (5.0/donor/OPU) were found with oocytes retrieved from donors treated with FSH (STIMUFOL®, Belgium) at a dosage of 150 IU per donor, compared to two others commercial FSH sources. Experiment 2: There were no differences in embryo development or yield with STIMUFOL FSH (total FSH 150 IU/donor) at a priming duration of either 60-h (Regime 1, six FSH injections) or 36-h (Regime 2, four FSH injections). Experiment 3: Compacted COCs required 22-26-h maturation in vitro (IVM) before IVF for optimal blastocyst development (36.1-41.1 %); however, short (18-h) and prolonged (30-h) IVM duration resulted in lower embryonic development. In contrast, expanded COCs resulted in inferior blastocyst development compared to compacted COCs. Immunofluorescence microscopy revealed that the ratio of 89.8 % cumulus compacted COCs were at the germinal vesicle (pachytene) phase while 98.9 % cumulus expanded COCs went through spontaneous meiosis from meiotic metaphase I, anaphase I, telophase I to metaphase II upon OPU retrieval (P<0.05). Pregnancy rates were not different among three FSH sources or different FSH treatments as long as embryos reached the blastocyst stage. Our study found that different sources of FSH used for Wagyu donor priming prior to OPU resulted in differential embryo development potentials, but those embryos that reached out to blastocysts had a competent implantation ability.

Effects of donor age and reproductive history on developmental potential of ovum pickup oocytes in Japanese Black cattle (Wagyu).

The objectives of this study were to analyze the (1) effects of donor age and multiparity on development of in vitro fertilization (IVF) embryos after ovum pickup (OPU), (2) effects of repeated and consecutive OPU-IVF procedures on embryo development, and (3) embryo production from OPU-IVF in donors with differing embryo yields after multiple ovulation and embryo transfer technology (MOET) in Japanese Black cattle (Wagyu). Donors were pre-treated with low-dosage follicle-stimulating hormone (FSH; 200 IU total), and oocytes were collected via OPU and fertilized by IVF to generate blastocysts. The number of oocytes collected per OPU session per donor was lower in heifers (2-4 years old, 5.3 oocytes) than in primiparous and pluriparous cows (2-10 years old, 13.6-19.1 oocytes; P < 0.05). Rates of blastocyst development for oocytes from heifers (33.1%) were lower than for those from cows (2-10 years old, 44.1-54.3%; P < 0.05), and average blastocyst yield/OPU/animal was lower in heifers (3.7) than in 5-6 years old cows (10.1; P < 0.05). Donors undergoing five consecutive OPU-IVF sessions after low-dosage FSH showed similar oocyte retrieval (12.2-15.1 oocytes per OPU/animal), blastocyst development rates (35.6-45.0%), and embryo yield/OPU/animal (4.8-5.8; P > 0.05) across sessions. Additionally, embryo yield from OPU-IVF was significantly improved in animals with previous low embryo yield from MOET (5.9 vs. 2.6, respectively, P < 0.05). These results indicate that Wagyu cows with previous births can be more productive as OPU-IVF donors than heifers, and oocytes from donors undergoing to five consecutive OPU-IVF cycles are competent for embryo development without loss of embryo yield/OPU/animal. Moreover, OPU-IVF can be used for embryo production and breeding from all elite Japanese Black cattle, regardless of previous low embryo yield in routine MOET.

Flame-Retardant Thermoplastic Polyether Ester/Aluminum Butylmethylphosphinate/Phenolphthalein Composites with Enhanced Mechanical Properties and Antidripping.

Aluminum butylmethylphosphinate AiBMP as a flame retardant and phenolphthalein as a synergistic agent were applied in a thermoplastic polyester elastomer (TPEE)) in the current study. The thermal properties, flame retardancy, crystallization and mechanical properties of TPEE/AiMBP with or without phenolphthalein were investigated using various characterizations, including the limiting oxygen index (LOI), vertical burning test (UL 94), thermogravimetric analysis TG, differential scanning calorimetry, microcombustion calorimeter (MCC), scanning electron microscopy (SEM), and mechanical tests. The results revealed that AiBMP alone is an efficient flame retardant of TPEE. Adding 15 wt.% AiBMP increases the LOI value of TPEE from 20% to 36%. The formula TPEE-15 AiBMP passed the UL 94 V-0 rating with no dripping occurring. The MCC test shows that AiBMP depresses the heat release of TPEE. In comparison with pure TPEE, the heat release rate at peak temperature and the heat release capacity of TPEE-15AiBMP are reduced by 46.1% and 55.5%, respectively. With the phenolphthalein added, the formula TPEE/13AiBMP/2Ph shows a higher char yield at high temperatures (>600 °C), and the char layer is stronger and more condensed than TPEE-15AiBMP.The tensile strength and elongation at break values of TPEE-13AiBMP-2Ph are increased by 29.63% and 4.8% in comparison with TPEE-15AiBMP. The SEM morphology of the fracture surface of the sample shows that phenolphthalein acts as a plasticizer to improve the dispersion of AiBMP within the matrix. The good char charming ability of phenolphthalein itself and improved dispersion of AiBMP make the TPEE composites achieve both satisfying flame retardancy and high mechanical properties.

Effect of Hole on Oil Well Cement and Failure Mechanism: Application for Oil and Gas Wells.

Cementing is an important operation in the drilling process. During the cementing process, mud cake, fracturing, perforation, and so on will cause holes in the cement sheath. Thereby, the size effect will reduce the cement strength, which will seriously affect the cementing quality. Several hole types were drilled to study the mechanical properties and damage mechanism of oil well cement. The stress distribution and failure process of cement containing a hole were studied using RFPA2D software. The experimental and simulation results demonstrate that an internal hole has an obvious effect on the cement performance. The hole will change the cement bearing capacity and affect the fracture direction. Three crack types exist: tensile, shear, and far-field. Both 2 and 5 mm vertical eccentric holes can reduce the cement tensile stress. Furthermore, the specimen tensile strength decreases with an increase in the diameter of holes. Horizontal eccentric holes with diameters of 2 and 5 mm will increase the cement tensile strength. Among these, the sample L2-2 exhibits a long crack path, high energy consumption, and a remarkable enhancement effect.

Study on the Adsorption Behavior between an Imidazolium Ionic Liquid and Na-Montmorillonite.

Interactions between 1-butyl-3-methylimidazolium tetrafluoroborate (IL), an ionic liquid, and Na-montmorillonite (Na-MMT) were studied under different kinetic conditions to investigate the adsorption behavior of IL by Na-MMT. The adsorption of IL by Na-MMT was rapid, with a fast rate, reaching a capacity of 0.43 mmol/g, lower than Na-MMT's cation exchange capacity (CEC) of 0.90 mmol/g. Meanwhile, the highest adsorption rate occurred at the IL concentration of 1000 mg/L. The exchangeable cation of Na-MMT could not be completely substituted by the cation group of IL regardless of the IL concentration. Stoichiometric desorption experiments confirmed that the cation exchange was the dominating adsorption mechanism for the IL adsorption by Na-MMT. The pH value of the solution between 2 and 11 had a negligible effect on the adsorption amount of IL by Na-MMT. The cation group of IL interacted into the interlayer of Na-MMT successfully, resulting in the change in the wettability of Na-MMT. A bilayer formation of the cationic group should occur in the interlayer of the modified Na-MMT and the configuration of IL was dependent on the adsorption amount of IL. Furthermore, the thermal stability of the modified Na-MMT was also dependent on the adsorption amount of IL.

Akt regulates ß-catenin in a rat model of focal cerebral ischemia-reperfusion injury.

The present study aimed to investigate the effects of the phosphoinositide 3­kinase (PI3K)/Akt signaling pathway on the Wnt/ß­catenin signaling pathway in rats with focal cerebral ischemia­reperfusion injury. A total of 96 rat focal cerebral ischemia­reperfusion models, established according to a modified version of Longa's method, were randomly divided into four groups: Sham­operated (S), cerebral ischemia­reperfusion injury (I), cerebral ischemia­reperfusion + basic fibroblast growth factor (bFGF) post­processing and, finally, cerebral ischemia­reperfusion + bFGF post­processing + PI3K inhibitor LY294002 (LY). Each group consisted of 24 rats and each group was divided into four subgroups according to the indicated reperfusion times of 12, 24, 48 and 72 h. The morphological changes of the cortical tissue and the cellular apoptosis were determined using hematoxylin and eosin staining and the terminal deoxynucleotidyl transferase dUTP nick end labeling method, respectively. The expression levels of phosphorylated (p­)Akt, glycogen synthase kinase­3ß (GSK­3ß) mRNA and ß­catenin in the cortical tissue were detected at different time­points. The number of apoptotic cells and the expression levels of p­Akt, GSK­3ß mRNA and ß­catenin in the I and LY groups were significantly higher compared with those in the S group (P<0.05). In the bFGF group, the number of apoptotic cells and the mRNA expression levels of GSK­3ß were significantly decreased, whereas the expression levels of p­Akt and ß­catenin were significantly increased compared with those in the I and LY groups (P<0.05). In cerebral ischemia­reperfusion injury, the PI3K/Akt signaling pathway regulated ß­catenin, the main member of the Wnt signaling pathway, via GSK­3ß, providing information to assist in further investigation of the mechanism of ß­catenin in ischemia­reperfusion injury.