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.
This study was performed to optimize the preparation of frozen lotus roots. Prior to freezing, an optimal blanching condition at $100^{\circ}C$ for 5 min was established, based on the microbial growth, texture, total phenolic content (TPC), and sensory evaluation results. The blanched samples were then frozen under various freezing conditions ($-20^{\circ}C$ in a freezer for 2 hr, $-70^{\circ}C$ in a gas nitrogen convection chamber for 7 min, and $-196^{\circ}C$ in liquid nitrogen for 20 sec), and their qualities after thawing were determined. The scanning electron microscopic analysis indicated that the microstructure of the sample frozen at $-70^{\circ}C$ was similar to that of the control sample, compared with the other freezing conditions (-20 and $-196^{\circ}C$). The antioxidant activities of the frozen samples decreased compared to those of the control, but there was no significant (p<0.05) difference among the treatments. In terms of TPC, the samples frozen at -70 and $-196^{\circ}C$ had significantly (p<0.05) higher values than the sample frozen at $-20^{\circ}C$. In addition, the drip loss of the sample frozen at $-20^{\circ}C$ was higher than those of the other frozen samples. These results suggest that freezing at $-70^{\circ}C$ in a gas nitrogen convection chamber can be an optimal freezing method of producing high-quality frozen lotus roots.
Journal of the Korea Institute of Building Construction
/
v.15
no.3
/
pp.265-271
/
2015
This study is intended to produce a PC (Precast Concrete) member without a steam curing process in developing the high early strength concrete satisfying the condition of 10MPa in compressive strength at the age of 6 hours, and is intended to ensure economic feasibility by increasing the turnover rate of concrete form. Hence, high early strength cement with high $C_3S$ content and the hardening accelerator of powder type accelerating the hydration of $C_3S$ was used. And the properties of concrete were evaluated according to the hardening accelerator mixing ratio (0, 1.2, 1.6, 2.0). No big difference was found from the tests of both slump and air content. When 1.6 % or higher amounts of the hardening accelerator were mixed, the compressive strength of 10MPa was achieved at the age of 6 hours. From the test results of autogenous (drying) shrinkage and plastic shrinkage, it can be seen that there was a difference according to hydration reaction rate due to the addition of the hardening accelerator. However, it was shown that no problem arose with crack and durability. And it was shown that resistance to freezing-thawing, carbonation, and penetration were excellent.
Journal of the Korea institute for structural maintenance and inspection
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v.14
no.5
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pp.128-135
/
2010
Polymer-modified cement mortar(PCM) has been widely used for strengthening of the concrete structures due to its excellent physical properties such as high strength and durability. Adhesive strength or behavior, on the other hands, between PCM and concrete is very important in strengthening the concrete member using PCM. Therefore the adhesive failure mechanism between PCM and concrete should be fully verified and understood. This study was performed to evaluate adhesive strength of PCM to the concrete by the direct pull-out test. In the direct pull-out tests, the adhesive strength under the various pre-treatment conditions such as immersion, thunder shower, freezing and thawing are evaluated. Also, the field direct pull-out test are performed to investigate the adhesive strength of mock-up test specimens. In the results of the test, the adhesive strength value by field test are lower than those of the standard curing condition. From these comparison and investigation, field test result was similar with the thunder shower test result. The results of the test was used to evaluate the korean industrial standard of polymer modified cement mortars for maintenance in concrete.
Journal of The Geomorphological Association of Korea
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v.19
no.3
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pp.71-82
/
2012
This research was carried out to estimate annual erosion and retreat rates by using datum-point and to identify the characteristics and causes of seasonal variations of sea-cliff slope in Pado-ri, Taean-gun. In the result, the erosion and retreat rates of sea-cliff were increased from spring to summer. The rates were increased rapidly between August and October, caused by the effects of extreme weather events such as severe rainstorms and typhoons, etc. Since then, the erosion and retreat rates of sea-cliff were decreased gradually, but the rates were increased again in winter due to the storm surge and mechanical weathering resulting from the repeated freezing and thawing actions of bed rocks. The factors that affect erosion and retreat rates of sea-cliff include the number of days with antecedent participation and daily maximum wave height. In particular, it turned out that the erosion is accelerated by strong wave energy during storm surges and typhoons. The annual erosion and retreat rates of study area for the past two years(from May 2010 to May 2012) were approximately 44~60cm/yr in condition of differences in geomorphological and geological characteristics at each point. These erosion and retreat rates were found to be higher than results of previous researches. This is caused by coastal erosion forces strengthened by extreme weather events. The erosion and retreat process of sea-cliff in the study area is composed by denudation of onshore areas in addition to marine erosion(wave energy).
Ten historic sites (denoted as A- J in this study) of a tomb were found during the construction of the east-west expressway in District 2 of Hyangnam, Hwaseong, which is implemented by the Gyeonggi-do headquarters of the Korea Land & Housing Corporation. Thetombswere first detected at siteH, and further investigations revealed various tombs from the Three Kingdoms period; artifacts such as gilt-bronze shoes and caps were excavated from wooden coffins in the tombs. The pottery examined in this study was the only pottery artifact excavated from the site. Its raw clay was soft and loose, reddish brown, and had quartz and feldspar particles of < 1 mm, which appeared to have been added as reinforcing agents. The firing temperature of the pottery was estimated to be under 800-870 ℃ as the mica remains and tridimite, which is the phase transition mineral of quartz, was not produced; a slight endothermic peak was also detected because of the hydration of sericite at 800 ℃. The condition of the artifact was severely weakened because of various factors, such as soil pressure from the stratum formed over the site and repeated freezing and thawing. The artifact could not be collected alone, and thus, surrounding soil that had attached to the artifact was also collected; the artifact was transported to the laboratory and conservation treatment was conducted in a safe and systematic manner.
Journal of The Geomorphological Association of Korea
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v.17
no.4
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pp.71-83
/
2010
In this study, the volume of shoreline retreat at sea-cliffs in the Taean peninsula(West Coast of Korea) was estimated and their erosion and seasonal landforms characteristics changes were investigated through multi-temporal aerial photographs and field survey. Based on the analysis of aerial photographs through ortho-correction, the results show that the length of shoreline and erosion area increase as erosion at sea-cliffs occurs in Pado-li and Dundu-li. To obtain the seasonal quantitative landforms changes and retreat of sea-cliffs, we marked top, middle, and bottom datum-points, from which the distance to the nearest bedrock was repeatedly measured. In these regions, the retreat of sea-cliffs gradually increases in spring to summer, but gradually decreases in autumn. In particular, the typhoon that has a great influence on the Korean peninsula in July to September in summer would drastically increase the retreat of sea-cliffs in comparison with other seasons. As the outcrop of sea-cliffs repeats freezing and thawing in winter, the retreat of sea-cliffs increases a little due to active mechanical weathering. To know the erosion and seasonal landforms changes of sea-cliffs, we took pictures of them in every month and then analyze their condition. The retreat of sea-cliffs was repeatedly occurred by the circulation of the erosion of sea-cliff base, landslides, the formation of slope sediment debris and their erosion, in that order.
Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
Journal of Embryo Transfer
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v.19
no.2
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pp.155-163
/
2004
The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.
Choi, Su Jin;Lee, Sun Hee;Song, In Ok;Koong, Mi Kyoung;Kang, Inn Soo;Jun, Jin Hyun
Clinical and Experimental Reproductive Medicine
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v.33
no.4
/
pp.237-243
/
2006
Objective: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). Methods: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test Results: Mean ages were similar between fresh ET ($40.0{\pm}1.8$ years, n=206) and frozen-thawed ET ($39.9{\pm}1.9$ years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET ($31.2{\pm}2.3$ years, n=40) and frozen-thawed ET ($31.9{\pm}3.1$ years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). Conclusion: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.
Kim, Yoo-Gon;Lee, Woo-Jong;Won, Chan-Hee;Kim, Yong-Hee;Yun, Ji-Sun;Hong, Min-Seon;Shin, Chul-Soo
Analytical Science and Technology
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v.24
no.3
/
pp.193-199
/
2011
The purpose of this experiment is to evaluate the stability of products according to the storage methods, the period of use, and the diurnal variations through the short term stability experiment of recombinunt protein A (rProtein A) produced in AP Tech Co. That is, we investigated how long the stability of the products would last, when we used the samples frozen at $-20^{\circ}C$, which is one of the storage conditions of the produced rProtein A and then kept them refrigerated at $4^{\circ}C$. The experiment was conducted for 8 weeks and 6 experiment points were established. The experiment was done by thawing the samples frozen at $-20^{\circ}C$ at room temperature, and then refrigerating them at $4^{\circ}C$. In addition, experiments for endotoxin, bioburden, HPLC purity, and concentration were conducted. As a result of the experiment, 0.5 EU/mg endotoxin was detected both at the beginning and at the 8th week and bioburden was not analyzed. In the case of purity, it showed 99.23~99.90% at 210 nm (RSD% 0.23%) and 100% at 280 nm, which meant the change into other materials didn't happen and there was no material degradation characteristics. Finally, we also found the fact that the concentration stayed stable at 55.15 mg/mL (RSD% 0.55%) both at the beginning and at the end. From the experiment results, we were able to conclude that the stability at the condition to store rProtein A at 4 oC for 8 weeks was procured without producing microorganisms or having material degradation characteristics.
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