Slow freezing cryopreservation of Korean bovine blastocysts with an additional sucrose pre-equilibration step.

Introduction: Embryo cryopreservation is a valuable technique used for preserving genetic resources for long periods. However, the survival rate of embryos is dependent on the method used. Therefore, in this study, we evaluated the efficiency of slow freezing method but with an additional dehydration step prior to freezing to overcome the formation of ice crystals. Methods: Oocytes collected from the ovaries of native Korean cattle subjected to in vitro fertilization were cultured for 7 days until the formation of expanded blastocysts. Before freezing, the blastocysts were placed in four pre-equilibration media: a control medium with no addition of sucrose, and three experimental media with the addition of 0.1, 0.25, and 0.5 M sucrose, respectively. Then, the pre-equilibrated embryos were frozen. Embryo survival and hatching rates were evaluated morphologically at 24, 48, and 72 h after thawing. Immunofluorescence staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and gene expression analysis of the re-expanded blastocytes were examined 24 h after freeze-thawing. Results: The survival rate was significantly higher in the 0.1 M group than in the control group (p < 0.05), and the hatching rate at 72 h was significantly higher in the 0.25 and 0.5 M groups than in the control group (p < 0.05). TUNEL-positive cells were significantly lower in the 0.25 M group than in the control group (12.5 ± 0.9 vs. 8.3 ± 0.8; p < 0.05). The gene expression of BCL2 associated X, heat shock protein 70 kDa, and aquaporin 3 in the 0.25 M group was significantly lower than that in the control group (p < 0.05). Conclusion: Our study revealed that treatment with 0.25 M sucrose before slow freezing improved the viability of bovine embryos after freeze-thawing.

Correlations between Properties of Pore-Filling Ion Exchange Membranes and Performance of a Reverse Electrodialysis Stack for High Power Density.

The reverse electrodialysis (RED) stack-harnessing salinity gradient power mainly consists of ion exchange membranes (IEMs). Among the various types of IEMs used in RED stacks, pore-filling ion exchange membranes (PIEMs) have been considered promising IEMs to improve the power density of RED stacks. The compositions of PIEMs affect the electrical resistance and permselectivity of PIEMs; however, their effect on the performance of large RED stacks have not yet been considered. In this study, PIEMs of various compositions with respect to the RED stack were adopted to evaluate the performance of the RED stack according to stack size (electrode area: 5 × 5 cm2 vs. 15 × 15 cm2). By increasing the stack size, the gross power per membrane area decreased despite the increase in gross power on a single RED stack. The electrical resistance of the PIEMs was the most important factor for enhancing the power production of the RED stack. Moreover, power production was less sensitive to permselectivities over 90%. By increasing the RED stack size, the contributions of non-ohmic resistances were significantly increased. Thus, we determined that reducing the salinity gradients across PIEMs by ion transport increased the non-ohmic resistance of large RED stacks. These results will aid in designing pilot-scale RED stacks.

Amino Functionalization of Zirconium Diboride for High Dispersion Stability and Solid Loading.

Zirconium diboride (ZrB²) is an ultrahigh-temperature ceramic (UHTC) with excellent thermal and mechanical properties. To fabricate a complexly structured UHTC sintered body with high density a high-solid-loading slurry with low viscosity is required to fill the mold well. Extensive studies have been carried out to improve the dispersion properties, including those on solvents and surfactants. In this study, a surface treatment using polyethylenimine (PEI) was investigated. This reagent was used to provide ZrB² particles with desirable properties, such as electrostatic repulsive force and steric hindrance, by the functionalization of amine groups. Additionally, surface treatments with different concentrations of PEI were performed. The appropriate content for a high dispersity was determined. The surface functional groups of the PEI-treated ZrB² were observed by Fourier-transform infrared spectroscopy. The surface charge and enhancement in dispersion were evaluated by zeta potential and particle size distribution analysis, respectively. The rheological properties of the ZrB² slurries with various solid loadings were analyzed by viscosity measurements. In conclusion, ZrB² slurries with solid loadings up to 50 vol% could be prepared through the PEI treatments.

Holding of bovine blastocysts at suprazero temperatures using small molecules.

Although assisted reproductive technology (ART) currently exists, the only embryo preservation technology that is available is cryopreservation. In the present study, small molecules were used to hold embryos at room temperature. The basic medium for embryo holding for a short period of time at 4 °C, 10 °C and 20 °C consisted of 1% BSA non-cryopreservation medium (BNC) instead of fetal bovine serum. To maintain survival and prevent damage during embryo incubation, three candidate small molecules were selected-CHIR99021, Y-27632 and Thiazovivin-and their concentrations were optimized. The viability and hatching rate of embryos incubated at 10 °C were greater for Y-27632-BNC and CHIR99021+Y-27632-BNC compared to BNC. However, the rate was lower for Thiazovivin-BNC compared to BNC. Although there were no surviving embryos after incubation at 20 °C, the viability and hatching rate of embryos significantly increased in Y-27632-BNC and CHIR99021+Y-27632-BNC compared to BNC. The pregnancy rate of embryos incubated at 20 °C was also greater in the CHIR99021+Y-27632-BNC group compared to that in the frozen group. The mechanism by which small molecules enhance survival of embryos during incubation was investigated, and expression of heat shock protein 70 was observed to increase. The findings of this work may be useful in improving ART in the agricultural field.

Microarray analysis of embryo-derived bovine pluripotent cells: The vulnerable state of bovine embryonic stem cells.

Although there are many studies about pluripotent stem cells, little is known about pluripotent pathways and the difficulties of maintaining the pluripotency of bovine cells in vitro. Here, we investigated differently expressed genes (DEG) in bovine embryo-derived stem-like cells (eSLCs) from various origins to validate their distinct characteristics of pluripotency and differentiation. We identified core pluripotency markers and additional markers which were not determined as pluripotency markers yet in bovine eSLCs. Using the KEGG database, TGFß, WNT, and LIF signaling were related to the maintenance of pluripotency. In contrast, some DEGs related to the LIF pathway were down-regulated, suggesting that reactivation of the pathway may be required for the establishment of true bovine embryonic stem cells (ESCs). Interestingly, oncogenes were co-down-regulated, while tumor suppressor genes were co-up-regulated in eSLCs, implying that this pattern may induce abnormal teratomas. These data analyses of signaling pathways provide essential information on authentic ESCs in addition to providing evidence for pluripotency in bovine eSLCs.

Improvement of pregnancy rate by intrauterine administration of dexamethasone and recombinant human leukemia inhibitory factor at the time of embryo transfer in cattle.

Bovine embryos (day 5) were cultured to day 10 with or without 100 ng/mL PGF2α in medium supplemented with control; 100 nM Dex; 1,000 U/mL recombinant human leukemia inhibitory factor (rhLIF); or Dex+rhLIF. Although the rates to development to the blastocyst were not significantly different among groups, the hatching rate after additional culture with Dex +/or rhLIF was significantly higher in all supplemented groups than the control (p < 0.05). In the presence of PGF2α, the hatching rate was significantly restored in all supplemented groups relative to the group treated with only PGF2α and the control (p < 0.05). Embryo transfer (ET) was performed with blastocysts (day 7). PGF2α levels of control recipient cows were significantly higher in the circulatory blood samples collected 60 min after ET than in samples collected 60 min before ET (p < 0.005), and were decreased in cows injected with loading medium supplemented with Dex+rhLIF (p < 0.005). Pregnancy rate was significantly higher in the ET group that received supplemented embryo-loading medium than in the non-supplemented control (p < 0.05). The intrauterine administration of Dex and rhLIF at ET prevented increased PGF2α in circulatory blood and resulted in enhanced pregnancy rate.

Development of MgO-C Refractory Having High Oxidation Resistance by Metal Coating Process.

Graphite has been modified with a coating reagent, which can form coating layer of a metal on the surface of graphite, to effectively prevent the oxidation of graphite used as a carbon source, compared to common antioxidants. The oxidation of graphite is resisted by the high oxidation reactivity of metal and the oxygen barrier effect of coating layer. Therefore, the metal layer should be homogeneously and continuously coated on the surface of graphite and the coating efficiency of metal should be increased. The metal layer was formed with a metal precursor existed only with a ion phase in an aqueous solution. The unmodified graphite was totally degraded and oxidized after the combustion test at 1000 degrees C in air. However, as graphite was modified by the metal precursor, the color of carbon was not changed after the heat treatment. These results mean that the coating layer is individually and uniformly formed on a surface of graphite, delaying the oxidation of graphite. Consequently, MgO-C refractory with the high oxidation resistance could be successfully fabricated by the modification of graphite with the metal precursor.

Thiazovivin, a Rho kinase inhibitor, improves stemness maintenance of embryo-derived stem-like cells under chemically defined culture conditions in cattle.

Despite numerous reported attempts, successful isolation of genuine embryonic stem cells of cattle has been rare. Previous studies have shown that Thiazovivin, a Rho-associated kinase inhibitor, improves the survival and self-renewal of human embryonic stem cells. The present study demonstrates the effect of Thiazovivin on the derivation of embryo-derived stem-like cells. Attachment rates of blastocyst and embryonic cell clumps onto feeder cells in the Thiazovivin treatment group were greater than those of the control group. The pluripotency markers of the OCT4 and NANOG genes, and the adhesion molecule E-cadherin were increased by Thiazovivin treatment. This study suggests that Thiazovivin treatment improves the maintenance of stemness in a putative stem-like cell populations of cattle by promoting the expression of pluripotency marker genes, as well as enhancing the expression of the E-cadherin gene, resulting in an increase in cell adhesion.

In vitro culture of stem-like cells derived from somatic cell nuclear transfer bovine embryos of the Korean beef cattle species, HanWoo.

We established and maintained somatic cell nuclear transfer embryo-derived stem-like cells (SCNT-eSLCs) from the traditional Korean beef cattle species, HanWoo (Bos taurus coreanae). Each SCNT blastocyst was placed individually on a feeder layer with culture medium containing three inhibitors of differentiation (3i). Primary colonies formed after 2-3 days of culture and the intact colonies were passaged every 5-6 days. The cells in each colony showed embryonic stem cell-like morphologies with a distinct boundary and were positive to alkaline phosphatase staining. Immunofluorescence and reverse transcription-polymerase chain reaction analyses also confirmed that these colonies expressed pluripotent markers. The colonies were maintained over 50 passages for more than 270 days. The cells showed normal karyotypes consisting of 60 chromosomes at Passage 50. Embryoid bodies were formed by suspension culture to analyse in vitro differentiation capability. Marker genes representing the differentiation into three germ layers were expressed. Typical embryonal carcinoma was generated after injecting cells under the testis capsule of nude mice, suggesting that the cultured cells may also have the potential of in vivo differentiation. In conclusion, we generated eSLCs from SCNT bovine embryos, using a 3i system that sustained stemness, normal karyotype and pluripotency, which was confirmed by in vitro and in vivo differentiation.

Ceramic-Based Composite Solid Electrolyte for Lithium-Ion Batteries.

Solid-state electrolytes such as lithium ion conducting ceramic or solid polymers have been studied as substitutes for liquid electrolytes, but critical problems exist at the interfacial resistance between the solid electrode and the solid electrolyte. In this study, we combined 80 wt % Li1.3 Ti1.7 Al0.3 (PO4 )3 (LTAP) as a lithium ion conducting ceramic with 10 wt % of poly(vinylidene fluoride) (PVdF) as a binder, and 10 wt % 1 M LiPF6 in ethyl carbonate/dimethyl carbonate (EC/DMC) to develop a new composite electrolyte. The ceramic-based composite electrolyte showed thermal stability and high ionic conductivity through reduction of the interface resistance. The lithium ion conductivity of the composite electrolyte was 8.9×10-4  S cm-1 at room temperature without leakage. Electrochemical tests of the Li1+x Mn2 O4 -LTAP-based composite electrolyte-Li1-x Mn2 O4 cell showed that the composite could be utilized as a potential electrolyte for high-safety lithium-ion batteries.

Thermoelastic characteristics of thermal barrier coatings with layer thickness and edge conditions through mathematical analysis.

The thermoelastic behaviors of such as temperature distribution, displacements, and stresses in thermal barrier coatings (TBCs) are seriously influenced by top coat thickness and edge conditions, which were investigated based on the thermal and mechanical properties of plasma-sprayed TBCs. A couple of governing partial differential equations were derived based on the thermoelastic theory. Since the governing equations are too involved to solve analytically, a finite volume method was developed to obtain approximations. The thermoelastic characteristics of TBCs with the various thicknesses and microstructures were estimated through mathematical approaches with different edge conditions. The results demonstrated that the top coat thickness and the edge condition in theoretical analysis are crucial factors to be considered in controlling the thermoelastic characteristics of plasma-sprayed TBCs.

Adhesion phenomena between particles according to the content of organic binder in core for thin-wall casting.

The content of organic binder in a core for thin-wall casting has been controlled to investigate the adhesion phenomena of inorganic binder between starting particles, as directly related to the mechanical and thermal properties of the core. The inorganic binder precursor was composed of tetraethyl orthosilicate and sodium methoxide as the silicon dioxide and sodium oxide precursors, respectively. Poly(vinyl alcohol), a hydrophilic polymer, was used as an organic binder. The particles coated with the inorganic precursor were sculpted with the organic binder and then the prepared core samples were heated at 1000 degrees C for 1 h. The core samples prepared with the optimum content of organic binder show the highest fracture strength. This may be due to the enhancement of adhesion by the glass phase formed between starting particles. However, when too much or too little organic binder is employed, the strength values of the core samples are significantly decreased. This is because the network structure of the glass phase is not inadequately created or the glass phase is not uniformly developed between starting particles, resulting in the insufficient contact between starting particles during the convert process.

Fabrication of nanosized Al2O3 coated reinforcing particle using electrostatic force.

Titanium carbide (TiC) particles have been coated with aluminum (Al) phase to enhance the compatibility with a metal matrix based on Al, expecting the homogenous dispersion of TiC particles into the matrix. The TiC particles were uniformly dispersed in the aqueous solution of pH 12. The aluminum nitrate as a precursor of Al phase was added to the solution dispersed with the TiC particles. The coating of Al phase onto the TiC particle was driven by the attractive force between the TiC particle with a negative charge and the Al cation in the aqueous solution. The TiC was not oxidized after heat treatment at 500 degrees C, whereas titanium dioxide peaks are detected after heat treatment at 1000 degrees C. In addition, the Al phase coated on the TiC surface was converted to amorphous alumina (Al2O3) and crystallized into alpha-Al2O3 during heat treatment at 500 and 1000 degrees C, respectively. The heat treatment condition such as atmosphere and temperature was an important factor in fabricating the reinforcing particles without degradation such as oxidation and phase transformation. The nano-sized Al2O3 coated reinforcing particles could be well fabricated using electrostatic force in the aqueous solution and by controlling the heat treatment condition.

Thermal Fatigue Behavior of Air-Plasma Sprayed Thermal Barrier Coating with Bond Coat Species in Cyclic Thermal Exposure.

The effects of the bond coat species on the delamination or fracture behavior in thermal barrier coatings (TBCs) was investigated using the yclic thermal fatigue and thermal-shock tests. The interface microstructures of each TBC showed a good condition without cracking or delamination after flame thermal fatigue (FTF) for 1429 cycles. The TBC with the bond coat prepared by the air-plasma spray (APS) method showed a good condition at the interface between the top and bond coats after cyclic furnace thermal fatigue (CFTF) for 1429 cycles, whereas the TBCs with the bond coats prepared by the high-velocity oxygen fuel (HVOF) and low-pressure plasma spray (LPPS) methods showed a partial cracking (and/or delamination) and a delamination after 780 cycles, respectively. The TBCs with the bond coats prepared by the APS, HVOF and LPPS methods were fully delaminated (>50%) after 159, 36, and 46 cycles, respectively, during the thermal-shock tests. The TGO thickness in the TBCs was strongly dependent on the both exposure time and temperature difference tested. The hardness values were found to be increased only after the CFTF, and the TBC with the bond coat prepared by the APS showed the highest adhesive strength before and after the FTF.

Dominant factors governing the rate capability of a TiO2 nanotube anode for high power lithium ion batteries.

Titanium dioxide (TiO(2)) is one of the most promising anode materials for lithium ion batteries due to low cost and structural stability during Li insertion/extraction. However, its poor rate capability limits its practical use. Although various approaches have been explored to overcome this problem, previous reports have mainly focused on the enhancement of both the electronic conductivity and the kinetic associated with lithium in the composite film of active material/conducting agent/binder. Here, we systematically explore the effect of the contact resistance between a current collector and a composite film of active material/conducting agent/binder on the rate capability of a TiO(2)-based electrode. The vertically aligned TiO(2) nanotubes arrays, directly grown on the current collector, with sealed cap and unsealed cap, and conventional randomly oriented TiO(2) nanotubes electrodes were prepared for this study. The vertically aligned TiO(2) nanotubes array electrode with unsealed cap showed superior performance with six times higher capacity at 10 C rate compared to conventional randomly oriented TiO(2) nanotubes electrode with 10 wt % conducting agent. On the basis of the detailed experimental results and associated theoretical analysis, we demonstrate that the reduction of the contact resistance between electrode and current collector plays an important role in improving the electronic conductivity of the overall electrode system.

Non-Prestonian behavior of rectangular shaped ceria slurry in shallow trench isolation chemical mechanical planarization.

Rectangular ceria particles were synthesized using the flash creation method. The influence of the morphology of ceria particles and the surfactant concentration on the removal rate was systematically investigated. These ceria slurries with polymeric surfactant molecules as the passivation agents of Si3N4 film, shows an exceptional non-Prestonian behaviors. The non-Prestonian behavior can be attributed to the increase in the contact area of the ceria particles with the SiO2 film, which is dominated by the morphology of the ceria particles. Force measurements using an atomic force microscope (AFM) at different concentrations of polymeric surfactant molecules was used to identify the interactions between the polymeric molecules and the oxide film and analyze the non-Prestonian behavior of ceria slurry having rectangular abrasives.

Microstructure and mechanical properties of Al2O3 composites with surface-treated carbon nanotubes (CNTs): dispersibility of modified carbon nanotubes (CNTs) on Al2O3 matrix.

Aluminum oxide (Al2O3) matrix have been reinforced by the multi-walled carbon nanotubes (MWCNTs) to overcome the inherent brittleness of Al2O3 matrix. In order to increase mechanical properties of MWCNTs-Al2O3 composites, MWCNTs need to be well dispersed and individually incorporated in Al2O3 matrix. In this work, aluminum hydroxide (Al(OH)3) used as a Al2O3 precursor and MWCNTs were mixed in an aqueous solution for the homogeneous mixing of hetero-particles, as functions of the content of MWCNTs and the potential hydrogen (pH) of Al(OH)3 suspension. Firstly, MWCNTs were purified and modified by an acid reagent, inducing that the dispersibility of MWCNTs is increased in an aqueous solution by carboxylic group given on the surface of MWCNTs. The modified MWCNTs were added in the Al(OH)3 suspension, and then the mixture was filtered at room temperature. The filtered powders were formed using an uniaxial pressing and then densified by a pressureless heat treatment. As the pH is decreased the Al(OH)3 particles are well dispersed in an aqueous solution, due to the increment of repulsive force between particles with a same surface charge. MWCNTs are individually incorporated into Al2O3 matrix up to 1 vol.% MWCNTs, whereas MWCNTs are aggregated at the composite with 3 vol.% MWCNTs. Therefore, control of the pH and the MWCNTs content are key factors to be considered for the fabrication of MWCNTs-Al2O3 composites with high functional properties.

Mechanical properties of porous Al2O3 composite with surface modified multi-walled carbon nanotubes (MWCNTs).

Multi-walled carbon nanotubes (MWCNTs) have been reinforced in alumina (Al2O3) matrix to overcome the inherent brittleness of the Al2O3 matrix. In this work, MWCNTs were treated by acid to provide hydrophilicity to hydrophobic MWCNTs, inducing the homogeneous dispersion of MWCNTs in an aqueous solution. Aluminum hydroxide (Al(OH)3) as a Al2O3 precursor was added in the solution with the modified MWCNTs, and then this mixture solution was filtered at room temperature. The prepared powders were calcinated at 800-1000 degrees C to reduce the gas pocket in the matrix by decomposition of Al(OH)3. Then the calcinated powders were formed, and heat-treated. The porous MWCNTs-Al2O3 composites show higher mechanical properties in flexure strength and hardness than the porous Al2O3 without the reinforcement phase, which is attributed to the high mechanical properties of MWCNTs. However, higher MWCNTs contents in the composites decrease the mechanical properties due to the aggregation of MWCNTs in the composites. Therefore, control of the MWCNTs content and its dispersibility in the matrix are key factors to be considered for the fabrication of the porous MWCNT-Al2O3 composites.

Size effect in Ni-coated TiC particles for metal matrix composites.

Nickel (Ni) particles have been coated on the surface of titanium carbide (TiC) particles to enhance the dispersion of TiC particles into a molten metal and to achieve an improvement in the mechanical and thermal properties of the metal matrix. The adhesion of Ni particles on the surface of TiC particles is induced by the attractive force between the TiC with a negative charge and the Ni cation in an aqueous solution. The powders prepared with the relatively large particle sizes of 1, 4, and 40 microm show both TiC and Ni phases, whereas that prepared with a particle size of 0.02 microm shows complex phases of Ni, TiC, and TiO2 (titanium dioxide). The TiO2 phase is caused by the oxidation reaction between the TiC and oxygen. The 1 microm powder shows that the Ni is located only around the TiC without any self-aggregation and the TiC and Ni particles are isolated in the 4 and 40 microm powders, as confirmed in TEM images. The particle size is the essential factor in fabricating highly efficient Ni-coated TiC particles for metal matrix composites.

Fracture Modes in Curved Brittle Layers Subject to Concentrated Cyclic Loading in Liquid Environments.

Damage response of brittle curved structures subject to cyclic Hertzian indentation was investigated. Specimens were fabricated by bisecting cylindrical quartz glass hollow tubes. The resulting hemi-cylindrical glass shells were bonded internally and at the edges to polymeric supporting structures and loaded axially in water on the outer circumference with a spherical tungsten carbide indenter. Critical loads and number of cycles to initiate and propagate near-contact cone cracks and far-field flexure radial cracks to failure were recorded. Flat quartz glass plates on polymer substrates were tested as a control group. Our findings showed that cone cracks form at lower loads, and can propagate through the quartz layer to the quartz/polymer interface at lower number of cycles, in the curved specimens relative to their flat counterparts. Flexural radial cracks require a higher load to initiate in the curved specimens relative to flat structures. These radial cracks can propagate rapidly to the margins, the flat edges of the bisecting plane, under cyclic loading at relatively low loads, owing to mechanical fatigue and a greater spatial range of tensile stresses in curved structures.