Park, Bola;Lee, Kung-Jin;Lee, Sang-Hee;Lee, Eunsong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
Journal of Embryo Transfer
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v.28
no.3
/
pp.193-198
/
2013
This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide($H_2O_2$) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.
To evaluate the practical application of oyster shells(OS) as construction materials, an experimental study was performed. More specifically, the long-term mechanical properties and durability of concrete blended with oyster shells were investigated. Test results indicate that long-term strength of concrete blended with 10% oyster shells is almost identical to that of normal concrete. However, the long-term strength of concrete blended with 20% oyster shells is appreciably lower than that of normal concrete. Thereby, concrete with higher oyster shell blend has the possibility of negatively influencing the concrete long-term strength. Elastic modulus of concrete blended with crushed oyster shells decreases as the blending mixture rate increases. Namely, the modulus is reduced to approximately 10∼15% when oyster shells are blended up to 20% as the fine aggregate. The drying shrinkage strain increases with an increasing crushed oyster shells substitution rate. In addition, the existing model code of drying shrinkage and creep do not coincide with the test results of this study. An adequate prediction equation needs to be developed. The utilization of oyster shells as the fine aggregate in concrete has an insignificant effect on fleering and thawing resistance, carbonation and chemical attack of concrete. However, water permeability is considerably improved.
For exposed slopes, the weathering degree over time has a major effect on the engineering properties of rocks and the slope stability. Rocks are gradually changed by weathering into soil over time, and the resulting physical, chemical and mechanical changes of rocks affect the engineering stability of the slope. However, there are not many ways to objectively evaluate the weathering degree of a slope. In this study, therefore, to investigate the weathering characteristics of rocks, granite, gneiss and shale distributed in the Chungbuk region were sampled by weathering stage and changes in their component minerals and tissues were investigated. Furthermore, artificial weathering was induced using the freezing and thawing test and quantitatively investigated through porosity and absorption rate. In addition, the changes of microcracks due to artificial weathering were evaluated through box fractal dimension ($D_B$). Through mineralogical study the phase change of constituting minerals, the growth of secondary minerals, the development of micro-cracks and the fabric changes due to weathering were observed. The mineralogical, chemical and engineering evaluations of the weathering degree through the experimental results in this study are expected to be useful for analyzing the weathering characteristics and causes by rock type and for proposing a methodology to evaluate the degradation of physical properties comparatively and quantitatively.
This study focus on the surface deterioration diagnosis based on the material characteristics of Taean Dongmunri Maaesamjonbulipsang(rock-carved standing triad Buddha Statue) and the weathering environment analysis. Rock materials of the triad Buddha is coarse-grained biotite granite which is composed mainly of quartz, K-feldspar, plagioclase, biotite and muscovite. The triad Buddha was serious surface irregularity(13.5%), exfoliation(12.3%) and discoloration(46.6%), and the physical characteristics using ultrasonic velocity appeared fourth grade (0.59, highly weathered). Korean style wooden shelter of the triad Buddha(closed type) maintained high humidity environment (daily mean; 86.6%) than exterior environment, and appeared the possibility of deterioration by freezing-thawing because the temperature showed below zero temperatures in winter. The shelter was changed from closed-type to open-type to relieve the moisture problem recently. But the moisture problem is yet to be solved because surface water flowing along northern wayside is flowed into triad Buddha. Therefore, environmental control will need to stop inflow of water into triad Buddha.
Kim Yong-jun;Park Young-jae;Kim Byeong-jin;Yu Il-jeoung
Journal of Veterinary Clinics
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v.11
no.2
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pp.545-552
/
1994
This experiment was carried out to investigate the renditions to maintain good post-thaw motility and viability of canine spermatozoa when the semen was frozen using methanol. The semen from two male dogs which had been proven to be fertile in the previous one year was treated with different compositions of semen diluent and was frozen at different freezing temperatures, When canine semen was frozen at-2$0^{\circ}C$, -6$0^{\circ}C$, or -8$0^{\circ}C$, the spermatozoa frozen and stored at -2$0^{\circ}C$ showed very low post-thaw motility and viability from day 2 to 7 and showed no viability since day 15 after freezing. The spermatozoa frozen and stored at -6$0^{\circ}C$ or -8$0^{\circ}C$ showed higher post-thaw motility and viability on day 2, 1, 15 and 30 after freezing than that frozen and stored at-2$0^{\circ}C$(p<0.01), with no difference between two groups. Among different composition groups of the semen diluents of control(tris + egg yolk + glycerol), egg yolk-free, 히ycerol-free, and tris-free, Prior to freezing, the egg yolk-free diluent showed significantly love. motility and viability than the other diluents(p<0.05). On each thawing day (from day 2 to 15 after freezing), control group showed considerably higher motility and viability than the other groups(p<0.01). The canine spermatozoa frozen and stored at -6$0^{\circ}C$ and -8$0^{\circ}C$ showed gradual decrease of motility from day 2 to 30 after freezing and the spermatozoa of these two groups thawed on day 30 showed considerably love. motility than those thawed on day 2 after freezing, respectively(p<0.01). These results indicate that the freezing temperature of either -6$0^{\circ}C$ or -8$0^{\circ}C$ can be applicable to the freezing method using methanol and also all of the components of the semen diluent including cryoprotectant, buffer and cold-shock buffer are very important to maintain motility and viability of canine spermatozoa in the freezing and thawing procedure.
This study was conducted to evaluate whether \"Royal jelly\" (RJ) added to Tris-buffer dilute contributed to supporting post-thaw viability and longevity of frozen canine spermatozoa. Two Japanese spitzs (2 to 4 years of age) were used as a semen donor. Semen was collected by manual masturbation and separated into 3 fractions. Only the sperm-rich fraction having sperm motility of more than 70%, containing sperm concentration of 2~4$\times$10$^{8}$ cells/ml and having dead or abnormal spermatozoa of less than 15% was used for the experiment. Each ejaculated semen was centrifuged at 400 $\times$ g for 5 min and then diluted in a Tris-buffer supplemented with 20 ml egg yolk (Ext I), 4% glycero1 and 1% Equex STM Paste (Ext II) or g1ycero1, Equex STM paste and RJ of various concentrations (Ext II-RJ). After freezing and thawing, viability of spermatozoa in Ext II -RJ containing 1% RJ immediately after thawing (67.5$\pm$9.6) was significantly lower than that of Ext II , Ext II -RJ containing 0.01 or 0.1% RJ (77.5$\pm$12.5, 78.7$\pm$8.2 and 80.0$\pm$6.3). However, Ext II-RJ containing 0.1% RJ yielded higher viability than Ext II, Ext II-RJ containing 0.01% at or 1% 1 h after thawing (69.5$\pm$8.1 vs. 55.0$\pm$12.9, 57.5$\pm$9.6 and 41.5$\pm$12.6; P<0.05). At 1 h after thawing, the viability of spermatozoa thawed in 7$0^{\circ}C$ (68.8$\pm$12.5) was significantly higher than that of spermatozoa thawed in 38$^{\circ}C$ (48.8$\pm$16.3), although there was no difference in the viability between both groups immediately after thawing (77.5$\pm$9.6 and 81.3$\pm$8.1). Post-thaw viability and longevity of post-thaw spermatozoa in Ext II-RJ containing 0.1% RJ was higher in those in Ext II at 1 h (65.0$\pm$12.9 vs. 42.5$\pm$12.6), 2 h (52.5$\pm$12.6 vs. 27.5$\pm$17.1) and 3 h (40.0$\pm$14.1 vs. 20.0$\pm$12.1) after thawing. These results indicated that addition of 0.1% af to Tris-buffer enhanced post-thaw viability and longevity of canine spermatozoa and this additive can be used for increasing the possibility of collision between spermatozoa and ova during insemination.emination.
Cement concrete pavement has become more common in Korean highway systems. However, as its service period increases, there are some technical problems occurs and no clear solution is available primarily due to the lack of active researches. This research, hence, aims to develop a new mix proportion that may provide better strength and durability with extended service life. Based on a variety of literature reviews, the experimental variables were determined as unit cement content, S/a ratio and W/C ratio. From the experimental works, it is recommended to increase the unit cement content up to 375kg/$m^3$, 400kg/$m^3$ and 425kg/$m^3$. The target slump and air content were set 40mm and 5%, respectively. The maximum size of coarse aggregate was decided to be 25mm because of the easiness of supply in the field. The reduction of W/C ratio was necessarily required and decreased to 0.4 which was proven not to cause any mixing problem with the increased unit cement contents along with polycarbon-based high range water reducing agent. In addition, it was known that the S/a ratio could be reduced to 0.34. The lowered S/a might be possible because of the increased cement paste and hence increased cohesiveness and workability.
This study was carried out to investigate the best condition for in vitro and in vivo culture after freezing and thawing of nuclear transplant 2 cell embryos. When nuclear transplant embryos were submitted to electrofusion, the significantly higher fusion rates of 2 cell donor nuclei were achieved at the electric field strength of DC 1.5 kV/cm for 100 and $150{\mu}sec$, DC 2.0 kV/cm for 100 and $150{\mu}sec$ than DC 1.0 kV/cm for 100 and $150{\mu}sec$(p<0.01). The significantly higher fusion rates of 4 cell donor nuclei were achievecl at DC 2.0 kV/cm for 100 and $150{\mu}sec$ than DC 1.0 kV/cm for 100 and $150{\mu}sec$(p<0.01). The fusion rates in 8 cell donor nuclei were 94.2~99.3%. The developmental potency to blastocyst in 2 cell donor nuclei was significantly higher in DC 2.0 kV/cm for $150{\mu}sec$ treated group(p<0.01). The significantly higher developmental potency to blastocyst in 4 cell donor nuclei were achieved at the electric field strength of DC 2.0 kV/cm for $150{\mu}sec$ than DC 1.5 kV/cm for 100 and $150{\mu}sec$, DC 2.0 kV/cm for $100{\mu}sec$ treated group(p<0.01). The develop mental potency to blastocyst in 8 cell donor nuclei was significantly higher in DC 2.0 kV/cm for $100{\mu}sec$ treated group(p<0.01). The developmental potency to blastocyst after nuclear transplantation was significantly higher in 2 cell donor nuclei than in 8 cell donor nuclei(p<0.01). When the recovered embryos in normal morphology were cultured in vitro, there were no significant differences in the developmental potency to blastocyst between the freezing methods and the concentrations of cryoprotectant(p<0.01). The production rates of offspring after transfer of nuclear transplant embryos to recipient mouse were no significant difference in 2, 4 and 8 cell donor nuclei.
Journal of the Korea Academia-Industrial cooperation Society
/
v.17
no.4
/
pp.766-771
/
2016
This study examined the long-term durability of PC boxes, which was manufactured by low-carbon eco-friendly concrete using an alternative binder to cement and alternative fine aggregate to sand and microwave heat curing system to reduce the construction cost of a near-surface transit system. Based on the test results, the initial compressive strength of microwave heat cured concrete was higher than that of the steam cured concrete, but those were similar in the long-term age. In addition, there was no significant difference between the two curing conditions in the chemical resistance and the freeze-thawing resistance, and the chloride ion penetration level of the concrete cured by two methods was very low. Therefore, low-carbon eco-friendly concrete and microwave heat curing technology are expected to contribute to the economic construction of a near-surface transit system, and reduce carbon dioxide emissions and environmental impact.
This study was performed 1) to establish the optimal PCR condition of sex determination in Hanwoo IVM/IVF embryos, 2) to examine the sex determination and sex ratio to the developmental stages of Hanwoo IVM/IVF embryos by two-step PCR method. The sexing of bovine IVF embryos were accurately determined by PCR methods using Y chromosome specific DNA primer(BOV 97M, 141bp) and bovine specific DNA primer(216bp). The fregment size were shown at 141 and 216 base pairs(bp) in male, and 216 bp in female. Two-steps PCR method in which the samples were amplified by 15 cycles with Y chromosome specific DNA primer and then amplified by additional 30 cycles with bovine specific DNA primer was effective in the sexing of bovine IVF embryos. The zona-free embryos were more effective than zona-intact embryos in bovine IVF embryo sexing. The appearance of Y chromosome specific band was 45.2% in embryos treated with protease K and 53.3% in embryos treated with freezing and thawing repeatedly. The optimun volume of DNA for sexing of Hanwoo IVF embryos were 2 to 10 $\mu$1 in Zona-free embryos and 12 to 13 $\mu$1 in zona-intact embryos. The sexing rate of bovine IVF embryos by PCR was 96.0% and questionable rate not identified sex was 4.0%, respectively. Among the sexed embryos, the percentage of male and female was 49.7% and 46.3%, respectively, the sex ratio was 1: 1.1. The successful rate of embryo sexing was increased to the developmental stages.
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