• Journal of Embryo Transfer
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• v.19 no.2
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• pp.155-163
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• 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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.12
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• pp.2019-2027
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• 2013

In this study, we tried to develop a coating agent for the fresh-cut fruits used in cakes. First, the coating agent mixing ratios of sugar, pectin, sodium alginate, carrageenan, xanthan gum, vitamin C, and purified water were selected to be 55, 2, 2, 0.04, 0.1, 0.05, and 40.81% (w/w), respectively. In a freeze-thaw stability of the coating agent, the viscosity remained constant for 3 cycles of freezing and thawing repetition process, but showed a slightly decreasing trend in the 4th repetition process (P<0.05). On the other hand, the sugar content, pH, and chromaticity remained constant even in the 4th repetition process. Pineapple coated with the coating agent had smaller weight loss, hardness changes, and total bacteria distribution compared to the uncoated pineapple (P<0.05). In the chromaticity, both of the two pineapples experienced browning with increasing storage duration, as L value decreases and b value increases. However, when the color difference was compared, the progress of browning for the uncoated pineapple was faster than the coated pineapple. Also, the progress of browning at $4^{\circ}C$ was found to be slower than the progress of browning at $25^{\circ}C$. Therefore, the storage stability of the fresh-cut fruits could be improved by coating the fresh-cut fruits for cakes with the coating agent and storing at a low temperature, which would contribute to extending the shelf-life of cakes.

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.199-206
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• 2012

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol ($39.85%{\pm}11.41$) than in 5% glycerol treatment ($18.08%{\pm}1.61$). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol ($34.12%{\pm}11.02$) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.8 no.3
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• pp.157-165
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• 1975

This experiment was carried out to evaluate the sanitary quality of commercially frozen sea foods. One hundred and sixteen samples in six different items from several refrigeration plant in Busan city were examined from March to December in 1974. In addition, the changes in bacterial density through the process from thawing, round or semifilleted frozen alaska pollack to the finishing as frozen fillet blocks were observed. To evaluate the sanitary quality, sanitary indicative bacteria such as total coliform, fecal coliform, fecal streptococci and enterococci as well as plate counts were determined. From the results, the median value of fecal coliform MPN was 20 per 100 grams of the samples and that of enterococci was 790. The median value of plate counts was $2.2\times10^4$ per gram. The plate counts were not correlated with the number of sanitary indicative bacteria. The results suggest that enterococci could be used advantageously in preference to coliform organisms as indicative bacteria for the evaluation of sanitary quality of frozen sea foods. The plate counts at $20^{\circ}C$ of the samples were 14 times higher than that at $35^{\circ}C$. Geometric mean of total coliform MPN was 310 and that of enterococci was 143. Bacterial density was reduced by fleering. Morethan 50 percent for total coliform MPN and $35^{\circ}C$ plate counts, and about 35 percent for enterococci MPN and $20^{\circ}C$ plate counts were reduced under the contact freezing unit which was generally operated at $-40^{\circ}C$. About fifty-five percent of the samples were negative in fecal coliform test and 10 percent of those were exceeded $1.0\times10^5$ per gram in $35^{\circ}C$ plate counts.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.4
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• pp.237-243
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• 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.

• Journal of the Korean Geotechnical Society
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• v.16 no.1
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• pp.179-189
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• 2000

The purpose of this research is to complement the existing researches on landfill bottom liners behavior during the periods of freeze and thaw. Landfill-related researches have been typically focused on small-scale soil samples that are often compacted under conditions different from those used in the field. Although these tests have been invaluable in clarifying the problem of freeze and thaw, extending the results of such experimental studies to prototype landfills are questionable. In this investigation, the author utilized a large scale laboratory simulation allowing inclusion of the field depth of the cover systems, layered soil profiles, rainfall simulation, a cold climate and boundary conditions similar to those encountered in the landfill. The soil materials were stabilized soils (mixed clays, cements, and minerals) instead of clays. The bottom liners are made up of drainage layer (30 cm), stabilized layer (75 cm), and leach collection layer (60 cm). The stabilized layers are made up of supporting layer (45 cm) and low permeable layer (30 cm) - consisting of $P_A\; and\; P_B$ layer. As a results, depths of penetration increased by about 2~5 more centimeters at rainfall simulated designs than those at no rainfall simulated designs (that is design 3, design 5 and design 7) - it increased by about 20mm/day in the bottom liners and frost heaves also increased it by a few millimeters. Also, a few cracks appeared partly. According to these results, we can surmise that the compacted stabilized soil is more reliable than the compacted clay liners for construction of the landfill liners.

• Parasites, Hosts and Diseases
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• v.24 no.2
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• pp.187-193
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• 1986

Enzyme-linked immunosorbent assay(ELISA) using crude and affinity-purified antigens of adult worms of Paragonimus westermani was performed for infected cat sera with different worm burden, from preinfection to 18th week after infection. Crude antigen was used with supernatant of homogenated worms by freezing-thawing method, and the supernate was centrifuged for 1 hour at 10,000 rpm at $4^{\circ}C$. Affinity-purified antigen(antibody-bound antigen) was prepared from fractions(bound and unbound) of crude antigen by affinity chromatography on CNBr-activated sepharose 4B, and IgG as a ligand was prepared from paragonimiasis cat serum(6 months infected) obtained by ammonium sulfate ($40%{\sim}45%$ saturated) precipitation method. By SDS-PAGE, crude antigen showed 22 polypeptide fractions while purified antigen showed 4 fractions: 36, 400, 34, 700, 27, 600 and 11, 500 in molecular weights. All cats were divided into five groups($G_1-G_5$) by different worm burdens. The mean of recovered worms(${\pm}SD$) and the number of cats in each group are as follows: $G_1$, 2 worms(0) and 4 cats; $G_2$, 4.75 (${\pm}0.66$) and eight; $G_3$, 10.75(${\pm}1.92$) and four; $G_4$, 23.20(${\pm}3.43$) and five; $G_5$, 48(${\pm}12.63$) and five cats. The results were summarized as follows: 1. The antibody levels(OD value) increased by worm burden in $G_1$ to $G_4$ generally. However, individual antibody levels were not exactly related with worm burden in all groups, especially there was a wide difference in $G_4$ and $G_5$. These results suggested that the worm burden in $G_4$ (about $20{\sim}30$ worms) is enough to produce antibody maximum in cats of $2{\sim}3kg$ weight. 2. The antibody levels increased significantly(p<0.05) compared to control sera at the 3rd week in $G_1$ and $G_2$, at the 2nd week in $G_3$, and at the 1st week in $G_4$ and $G_5$. Especially in the 4th week, OD value increased more in $G_1$(p<0.01) and in $G_2$ to $G_5$(p<0.001). In the pattern of antibody levels by ELISA in each group, OD in $G_1$ increased to the 18th week continuously, in $G_2$ OD was maintained same after the 16th week, but in $G_3$ it decreased after the 16th week, and it was maintained same in $G_4$ and $G_5$ after the 14th week. 3. The antibody levels by ELISA with the affinity-purified antigen were higher than those with crude antigen in all groups generally. Especially, the difference of OD values between two antigens was larger from the 4th to the 10th week. In $G_1$ and $G_2$ OD with purified antigen was higher than that with crude one to the 18th week. It was also higher in $G_3$ than that with crude antigen to the 16th week and OD of $G_4$ and $G_5$ were higher before the 14th week than that with crude antigen, however became lower at the 16th week. Consequently, the antibody level in ELISA with affinity-purified antigen was more sensitive at the early weeks after infection and in light infection groups than that with crude antigen.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.379-384
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• 1996

This study was undertaken to investigate the sister chromatid exchange (SCE) frequency and embryonic development after exposure to cryoprotectants and vitrification of mouse zygotes. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). After mouse zygotes were exposed to EFS40 at $25^{\circ}C$ for 30 sec., they were immediately plunged into $LN_2$ or cultured for cryoprotectant toxicity test without freezing. The results obtained in these experiments were summarized as follows: After thawing, survival rates to the 2-cell stage of zygotes exposed to or vitrified in EFS 40 (98.5%, 95.2%) were not significantly difference compared with that of control (100%). However, the developmental rates upto blastocyst and hatching blastocyst in vitrified groups (66.7, 50.0%) were lower than those of control (93.9, 81.8%) or exposed group (94.0, 78.8%) (p<0.05). When the influence of vitrification and exposure to cryoprotectant on the in vitro development of mouse zygotes was assessed by the SCE frequency, the SCE frequency in exposed ($20.2{\pm}2.1$) to or vitrified embryos ($21.4{\pm}3.2$) was higher than that in control embryos ($16.8{\pm}1.5$). These results suggest that the frequency of SCE was increased after cryoprotectant exposure or Vitrification although developmental rates of zygotes upto blastocysts and /or hatching blastocysts were not afected by cryoprotectant.

• Journal of the East Asian Society of Dietary Life
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• v.21 no.3
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• pp.345-352
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• 2011

TMR feed was developed by adding mugwort (Artemisia capillaris), and was fed to Hanwoo cattle to investigate the effects of feeding mugwort on the physicochemical and sensory characteristics of rump meat, and to determine the feasibility of producing Hanwoo beef with high quality and functionality. The experimental samples consisted of the Hanwoo rump from cattle fed with fattening TMR feed without mugwort (T0), and those fed with fattening cattle TMR feed supplemented with mugwort (T1). T1 was significantly higher than T0 for Hanwoo rump characteristics of Hunter's $L^*$, $a^*$, $b^*$ values (p<0.05). VBN content for T0 was significantly higher than for T1, and EDA for T1 was significantly higher than for T0 (p<0.05). There was no significant difference between T0 and T1 in terms of pH, TBARS, and total bacterial numbers. Water holding capacity for T1 was significantly higher than for T0 (p<0.05), but there was no significant difference between T0 and T1 in terms of freezing loss, thawing loss, and cooking loss. Springiness for T1 was significantly higher than for T0 (p<0.05), and there was no significant difference between T0 and T1 in terms of hardness, cohesiveness, gumminess, chewiness, and shear force. There was no significant difference between T0 and T1 in terms of acid value, peroxide value, and iodine value. However, the melting point for T1 was significantly lower than for T0 (p<0.05). Aroma of raw meat for T1 was significantly superior to aroma for T0 (p<0.05). Taste, palatability of boiled meat, and juiciness of roasted meat for T1 were significantly superior to those parameters for T0 (p<0.05). These results suggest that the feed containing mugwort can be used to improve color and sensory characteristics, inhibit VBN formation, and also to increase antioxidant ability as a functional feed.

• Journal of Food Hygiene and Safety
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• v.27 no.4
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• pp.420-426
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• 2012

The purpose of this study was to identify control points through microbiological hazard analysis in the manufacturing processes of starch noodles. Samples were collected from the ingredients, manufacturing processes, equipment and environment. Microbiological hazard assessments were performed using aerobic plate counts (APC), Enterobacteriaceae (EB), E. coli and five pathogens including B. cereus, E. coli O157:H7, L. monocytogenes, Salmonella spp., and S. aureus. The APC levels in raw materials were from 2.12 to 3.83 log CFU/g. The contamination levels after kneading were 4.31 log CFU/g for APCs and 2.88 log CFU/g for EB counts. APCs decreased to 1.63 log CFU/g and EB were not detected after gelatinization, but their levels slightly increased upon cooling, cutting, ripening, freezing, thawing, and separating. The reuse of cooling and coating water would be a critical source of microbial increase after cooling. After drying, APCs and EB counts decreased to 5.05 log CFU/g and 2.74 log CFU/g, respectively, and the levels were maintained to final products. These results suggest that the cooling process is a critical control point for microbiological safety, and the cooling water should be treated and controlled to prevent cross contamination by pre-requisite program.