Reactive oxygen species (ROS) production and F-actin cytoskeleton dynamics play important roles in the survival rate of blastocysts after the vitrifiedwarming process. However, the protective effects of Mito-TEMPO against cryo-injury and viability through F-actin aggregation and mitochondrial-specific ROS production in vitrificated-warmed bovine embryos have not been investigated. The aims of the present study were to: (1) determine the effects of Mito-TEMPO on embryonic developmental competence and quality by F-actin stabilization during in vitro culturing (IVC), and (2) confirm the effects of Mito-TEMPO through F-actin structure on the cryotolerance of vitrification-warming in Mito-TEMPO exposed in vitro production (IVP) of bovine blastocysts. Bovine zygotes were cultured with 0.1 μM Mito-TEMPO treatment for 2 days of IVC. Mito-TEMPO (0.1 μM) exposed bovine embryos slightly improved in blastocyst developmental rates compared to the non-treated group. Moreover, the viability of vitrified-warmed blastocysts from Mito-TEMPO treated embryos significantly increased (p < 0.05, non-treated group: 66.7 ± 3.2% vs Mito-TEMPO treated group: 79.2 ± 5.9%; re-expanded at 24 hours). Mito-TEMPO exposed embryos strengthened the F-actin structure and arrangement in the blastocyst after vitrification-warming. Furthermore, the addition of Mito-TEMPO into the IVC medium enhanced embryonic survival and quality through F-actin stabilization after the vitrification-warming procedure. Overall, our results suggest that supplementing the culture with 0.1 μM Mito- TEMPO improves the embryonic quality and cryo-survival of IVP bovine blastocysts.
Vitrification methods are commonly used for mammalian reproduction through the long-term storage of blastocyst produced in vitro. However, the survival and quality of embryos following vitrification are significantly low compared with blastocyst from in vitro production (IVP). This study evaluates that the survival of frozen-thawed bovine embryos was relevant to mitochondrial superoxide derived mitochondrial activity. Here we present supplementation of the cryopreservation medium with Mito- TEMPO (0.1 μM) induced a significant (p < 0.001; non-treated group: 56.8 ± 8.7%, reexpanded at 24 h vs Mito-TEMPO treated group: 77.5 ± 8.9%, re-expanded at 24 h) improvement in survival rate of cryopreserved-thawed bovine blastocyst. To confirm the quality of vitrified blastocyst after thawing, DNA fragmentation of survived embryos was examined by TUNEL assay. As a result, TUNEL positive cells rates of frozenthawed embryos were lower in the Mito-TEMPO treated group (4.2 ± 1.4%) than the non-treated group (7.1 ± 3.5%). In addition, we investigated the intracellular ROS and mitochondrial specific superoxide production using DCF-DA and Mito-SOX staining in survived bovine embryos following vitrification depending on Mito-TEMPO treatment. As expected, intracellular ROS levels and superoxide production of vitrified blastocysts after cryopreservation were significantly reduced (p < 0.05) according to Mito-TEMPO supplement in freezing medium. Also, mitochondrial activity measured by MitoTracker Orange staining increased in the frozen-thawed embryos with Mito-TEMPO compared with non-treated group. These results indicate that the treatment of Mito-TEMPO during cryopreservation might induce reduction in DNA fragmentation and apoptosis-related ROS production, consequently increasing mitochondrial activation for developmental capacity of frozen-thawed embryos.
The IMO (International Maritime Organization) has mandated the restriction of SOx emissions to 0.5 % for all international sailing vessels since January 2020. And, a number of countries have designated emission control areas for stricter environmental regulations. Three representative methods have been suggested to cope with these regulations; using low-sulphur oil, installing a scrubber, or using LNG (Liquefied Natural Gas) as fuel. In this paper, economic analysis was performed by comparing the method of installing a scrubber with the method of using low-sulphur oil without installing additional equipment. We suggested plausible layouts and compared the pros and cons of different scrubber types for retrofitting. We selected an international sailing ship as the target vessel and estimated payback time and benefits based on navigation route, fuel consumption, and installation and operation costs. Two case of oil prices were analyzed considering the uncertainty of fuel oil price fluctuation. We found that the expected payback time of investment varies from 1 year to 3.5 years depending on the operation ratio of emission control areas and the fuel oil price change.
We studied growth and physiological responses of Ambrosia trifida L. under the different light condition. Ambrosia trifida L. is Korean invasive species. For study, we selected the two spot (T1 and T2) in the Kongju National University. In each spots, we investigated growth and physiological responses in the more light environment (L) and in the less light environment (S). it makes 4 research spot (T1-L, T1-S, T2-L and T2-S). and first, we estimate light intensity (Lux) in each research spots. then take data on the growth responses which are a shoot height, a number of leaves, a leaf width and a leaf length. Also, we get some physiological data which are a photosynthetic rate, a transpiration rate, a stomata conductance, water use efficiency, chlorophyll content, photochemical efficiency of photosystem II (Fv/Fm). And, analysed them by using one-way ANOVA. As a results, the shoot height, No. of leaves, Leaf width and Leaf length were recorded relatively high in the spot of the most light quantity and in the spot of the least light quantity. In the physiological responses, the more light quantity was increased, the more photosynthetic rate, transpiration rate and stomatal conductance were increased. Fv/Fm, the photochemical efficiency of photosystem II, was the least recorded in the spot of the most light quantity. These results show Ambrosia trifida grow well in the spot of the most and the least light quantity but it grown in the shade has the less environmental stress.l
The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas9) system can be applied to produce transgenic pigs. Therefore, we applied CRISPR/Cas9 system to generate FoxN1-targeted pig parthenogenetic embryos. Using single guided RNA targeted to pig FoxN1 genes was injected into cytoplasm of in vitro matured oocyte before electrical activation. In results, regardless of the concentrations of vector, the cleavage rate were significantly (p<0.05) decreased (4 ng/μl, 51.24%; 8 ng/μl, 40.88%; and 16 ng/μl; 45.22%) compared to no injection group (70.44%). The blastocyst formation rates were also decreased in vector injected 3 groups (4 ng/μl, 7.96%; 8 ng/μl, 6.4%; and 16 ng/μl; 9.04%) compared to no injection group (29.07%). In addition, the blastocyst formation rates between sham injected group (13.51%) and no injection group (29.07%) also showed significant difference (p<0.05). The mutation rates were comparable between groups (4 ng/μl, 18.4%; 8 ng/μl, 12.5%; and 16 ng/μl; 20.0%). The sequencing analysis showed that blastocysts derived from each group were successfully mutated in FoxN1 loci regardless of the vector concentrations. However, the deletion patterns were higher than the patterns of point mutation and insertion regardless of the vector concentrations. In conclusion, we described that cytoplasmic microinjection of FoxN1-targeted CRISPR/Cas9 vector could efficiently generate transgenic pig parthenogenetic embryos in one-step.
Herein, we describe the effect of the cooling-off condition of a solution-processed 6,13-bis(triisopropylsilylethynyl)-pentacene (TIPS-pentacene) film on its molecular distribution and the resultant electrical properties. Since the solvent in a TIPS-pentacene droplet gradually evaporates from the rim to the center exhibiting a radial form of solute, for a quenched case, domains of the TIPS-pentacene film are aboriginally spread showing original features of radial shape due to suppressed molecular rearrangement during the momentary cooling period. For the slowly cooled case, however, TIPS-pentacene molecules are randomly rearranged during the long cooling period. As a result, in the lopsided electrodes structure proposed in this work, the charge transport generates more effectively under the case for radial distribution induced by the quenching technique. It was found that the molecular redistribution during the cooling-period plays an important role on the magnitude of the mobility in a solution-processed organic transistor. This work provides at least a scientific basis between the molecular distribution and electrical properties in solution-processed organic devices.
We analysed interfacial traps in organic thin-film transistors (TFTs) in which pentacene and 6,13-bis(triisopropylsilylethynyl)-pentacene (TIPS-pentacene) organic semiconductors were deposited by means of vacuum-thermal evaporation and drop-coating methods, respectively. The thermally-deposited pentacene film consists of dentritic grains with the average grain size of around 1 ?m, while plate-like crystals over a few hundred microns are observed in the solution-processed TIPS-pentacene film. From the transfer characteristics of both TFTs, lower subthreshold slope of 1.02 V/decade was obtained in the TIPS-pentacene TFT, compared to that (2.63 V/decade) of the pentacene transistor. The interfacial trap density values calculated from the subthreshold slope are about 3.4×1012/cm2 and 9.4×1012/cm2 for the TIPS-pentacene and pentacene TFTs, respectively. Herein, lower subthreshold slope and less interfacial traps in TIPS-pentacene TFTs are attributed to less domain boundaries in the solution-processed TIPS-pentacene film.
The pretreatment for substrate was carried out in change of gun pressure of bar using wet blasting. The size of powder was about . As the results, the surface roughness of cemented carbide substrate was improved with increment of gun pressure of wet blasting. A new surface layer was formed and Co particles were uniformly distributed over the entire surface after pretreatment. The adhesion of the pretreated substrate in same PVD-TiAlN film was improved and in approximately shown the best adhesion value.
A three-dimensional (3-D) method of analysis is presented for determining the free vibration frequencies and mode shapes of solid paraboloidal and complete (that is, without a top opening) paraboloidal shells of revolution with variable wall thickness. Unlike conventional shell theories, which are mathematically two-dimensional (2-D), the present method is based upon the 3-D dynamic equations of elasticity. The ends of the shell may be free or may be subjected to any degree of constraint. Displacement components ur, uθ, and uz in the radial, circumferential, and axial directions, respectively, are taken to be sinusoidal in time, periodic in θ, and algebraic polynomials in the r and z directions. Potential (strain) and kinetic energies of the paraboloidal shells of revolution are formulated, and the Ritz method is used to solve the eigenvalue problem, thus yielding upper bound values of the frequencies by minimizing the frequencies. As the degree of the polynomials is increased, frequencies converge to the exact values. Convergence to four digit exactitude is demonstrated for the first five frequencies of the complete, shallow and deep paraboloidal shells of revolution with variable thickness. Numerical results are presented for a variety of paraboloidal shells having uniform or variable thickness, and being either shallow or deep. Frequencies for five solid paraboloids of different depth are also given. Comparisons are made between the frequencies from the present 3-D Ritz method and a 2-D thin shell theory.
An exact solution procedure is formulated for the free vibration and buckling analysis of rectangular plates having two opposite edges simply supported when these edges are subjected to linearly varying normal stresses. The other two edges may be clamped, simply supported or free, or they may be elastically supported. The transverse displacement (w) is assumed as sinusoidal in the direction of loading (x), and a power series is assumed in the lateral (y) direction (i.e., the method of Frobenius). Applying the boundary conditions yields the eigenvalue problem of finding the roots of a fourth order characteristic determinant. Care must be exercised to obtain adequate convergence for accurate vibration frequencies and buckling loads, as is demonstrated by two convergence tables. Some interesting and useful results for vibration frequencies and buckling loads, and their mode shapes, are presented for a variety of edge conditions and in-plane loadings, especially pure in-plane moments.
Purpose – The structural changes of Korean agriculture are complex due to heterogeneous production processes and farms’ features. This study analyzed trends of dualism in Korean agriculture over the period 2000-15 based on farm-level data to clarify the specific trends of dualism in terms of farm income, farm-size, and farm operators’ age. From the results of this study, we would be able to understand the features of structural changes in Korean agriculture more profoundly. Research design, data, and methodology – We incorporated farm-level data in South Korea: Agricultural census and Farm household economy survey. As measures of inequality, we used size-weighted quantiles, and normalized Gini coefficients as well as mean and conventional quantiles. The size-weighted quantiles are more robust to changes in the number of small farms, but they are more sensitive to changes in the distribution of farm-size. Thus, they would be more useful to identify trends of dualism of Korean agriculture. Results - The results show that the farmland distribution of crop farms became more skewed and dispersed. However, the herd distribution of livestock farms became more concentrated. To be specific, their mean and 1st quantile increases more rapidly than their size-weighted 2nd quantile and size-weighted 3rd quantile. Gini coefficients of livestock farms regarding their herd distribution decreased by 0.1 on average. In the case of income distribution, the results indicate that the polarization regarding farm household/agricultural/non-agricultural income became more severe. However, we also found that the distribution of transfer income became concentrated continuously. The results imply that transfer income including subsidies would decrease farm income polarization. Lastly, during the study periods, Korean farms were aging over time, and age distribution of them more concentrated. Conclusions – The structure of Korean agriculture has been changing, even though the absolute size of it decreased over time. Land (herd) distribution became more dispersed (concentrated). Inequality regarding agricultural income became more severe, and it made farm household income more polarized even though transfer income would decrease income gaps among farms. Lastly, farms continue to age regardless of farm types and this might affect the structural changes in Korean agriculture in the future.