• Food Science of Animal Resources
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• v.29 no.4
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• pp.437-444
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• 2009

${\gamma}-Aminobutyric$ acid (GABA) producing lactic acid bacteria, Lactobacillus acidophilus RMK567 was cultivated in 50 L of sterilized MRS broth using a fermenter at $40^{\circ}C$ for 24 h. The cell number was increased to $10.04{\pm}0.13$ Log CFU/mL with a growth rate constant (k) of 0.454 generation/h and a generation time (g) of 2.303 h after a lapse of a lag phase (L) of 5.16 h. A total of 487 g of cell paste with 40.5% moisture was harvested with viable cell number of 12.48 Log CFU/g cell paste. The cell pastes after preparation with glycerol, glucose, and polydextrose as cryo-protectants were lyophilized under a vacuum of 84 m torr. A total of 408 g of freeze dried (FD) cell powders were mixed with a commercial strain of Streptococcus thermophilus to prepare of three types FD starter cultures with the viable cell numbers of 12.42 (FDA-GY), 12.60 (FDBGG) and 12.91 (FDC-GP) Log CFU/g. During preservation the FD cultures at -$18^{\circ}C$, the cell viability of the FD starter cultures were rapidly dropped to below 3.24% of the day of storage. No significant difference was found in the cell viabilities among three types of FD starters cultures, but significant difference (p<0.01) was found in storage periods. Yoghurts fermented through FD starter culture of L. acidophilus RMK567 were determined to contain $155.16{\pm}8.53$ ppm, $243.82{\pm}4.27$ ppm, and $198.64{\pm}23.46$ ppm of GABA, respectively. This study shows that GABA production activity of L. acidophilus RMK567 is not affected during the freeze drying process and would be available for commercial production of yoghurt containing high GABA content.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.2
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• pp.136-140
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• 1986

The change of cell counts of Vibrio parahaemolyticus in fish muscle by the storage time and temperature was examined to get basic informations for precautionary steps against food poisoning of slices of raw fish (sashimi). There fore, we inoculated fish homogenate of oceanic bonito (Katsuwonus pelamis), yellow tail (Seriola quinqueradiata) with Kanagawa positive Vibrio parahaemolyticus and stored it at $30^{\circ}C,\;18^{\circ}C,\;4^{\circ}C\;and\;-20^{\circ}C$ for 24 hours. The number of the Vibrio parahaemolyticus upon fish homogenate stored at $30^{\circ}C\;and\;18^{\circ}C$ decreased for the first two hours and increased thereafter. When the fish homogenates inoculated with Vibrio parahaemolyticus at about $10^3$ per gram were stored at $18^{\circ}C\;and\;30^{\circ}C$ for 10 hours, the cell numbers increased about 10 times and 1,000 times initial cell numbers, respectively. The survival rate of Vibrio parahaemolyticus was about $20\%$, when the inoculated fish homogenates were stored at $-20^{\circ}C$ for 24 hours. Vibrio parahaemolyticus inoculated in fish homogenates was decreased by about $10\%$ of initial cell numbers by the storage at $4^{\circ}C$ for 4 hours and it was decreased by about $50\%$ after 24 hours storage of the samples at the same temperature. The decreasing rate of inoculated Vibrio parahaemolyticus in fresh fish muscle homogenate was higher than that in frozen fish muscle homogenate during the storage time at a refrigerator.

• Korean Journal of Animal Reproduction
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• v.11 no.3
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• pp.161-169
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• 1987

This experiment was carried out to investigate the optimal cooling rate and the plunging temperature into liquid nitrogen of the 8-cell hamster embryos. The female hamsters were induced to superovulate by intraperitoneal injection of 30 i.u. PMSG. Embryos were flushed from oviduct and uterine horn. Embryos were frozen and incubated with a modified Dulbecco's phosphate buffered saline, and equilibrated with 1.5M-dimethyl sulfoxide by a 3-step procedure. The cooling rate of samples was 1$^{\circ}C$/min from room temperature to -5$^{\circ}C$ and the samples were seeded at -5$^{\circ}C$. The plunging temperatures into liquid nitrogen were -20, -25, -30, -35, -40, -45, -50 and -55$^{\circ}C$ at 0.3$^{\circ}C$/min, 0.5$^{\circ}C$/min and 1$^{\circ}C$/min cooling rates, respectively. This mean numbers of ovulation points and recovered embryos were 59.4 and 48.4 appearing 81.6% recovery rate. The percentage of 8-cell embryos in recovered embryos was 68.2. The survival rates of embryos plunged at -45$^{\circ}C$ were 73.5% at 0.3$^{\circ}C$/min, 44.8% at 0.5$^{\circ}C$/min and 30.3% at 1$^{\circ}C$/min cooling rates, respectively. This mean numbers of ovulation points and recovered embryos were 59.4 and 48.4 appearing 81.6% recovery rate. The percentage of 8-cell embryos in recovered embryos was 68.2. The survival rates of embryos plunged at -45$^{\circ}C$ were 73.5% at 0.3$^{\circ}C$/min, 44.8% at 0.5$^{\circ}C$/min and 30.3% at 1$^{\circ}C$/min cooling rates, respectively. The survival rates at 0.3$^{\circ}C$/min were significantly high. Under the condition of 0.3$^{\circ}C$/min cooling rate, the survival rates of embryos according to the plunging temperature were 70.0% and 73.5% at -40 and -45$^{\circ}C$, and those were higher than other plunging temperatures. Under the condition of 0.5$^{\circ}C$/min and 1$^{\circ}C$/min cooling rates, the survival rates according to the plunging temperatures were lower than the cooling rate of 0.3$^{\circ}C$/min, showing the similar tendency at all the plunging temperatures. In conclusion, 8-cell hamster embryos showed the best survival rates at 0.3$^{\circ}C$/min cooling rate and -45$^{\circ}C$ plunging temperature.

• Molecular & Cellular Toxicology
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• v.3 no.3
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• pp.198-207
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• 2007

[ $^{18}F$ ]-fluorodeoxyglucose (FDG) uptake on positron emission tomography (PET) scan has been found to reflect tumor aggressiveness and prognosis in various types of cancer. In this study, the gene expression profiles of glial tumors were evaluated to determine whether glial tumors with high $^{18}F$-FDG uptake have more aggressive biological potential than with low uptake. Surgical specimens were obtained from the 12 patients with glial tumors (4 males and 8 females, age range 42-68 years). The tumor samples were divided into two groups based on the $^{18}F$-FDG uptake PET scan findings: high $^{18}F$-FDG uptake (n=4) and low $^{18}F$-FDG uptake (n=8). The pathological tumor grade was closely correlated with the $^{18}F$-FDG uptake pattern: Glial tumors with high $^{18}F$-FDG uptake were pathologically Edmondson-Steiner grade III, while those with low uptake were grade II. The total RNA was extracted from the frozen tissues of all glial tumors (n=12), and adjacent non-cancerous tissue (n=3). The gene expression profiles were evaluated using cDNA microarray. The glial tumors with high $^{18}F$-FDG uptake showed increase expression of 15 genes compared to those with low uptake (P<0.005). Nine genes were down-regulated. Gene expression is closely related to cell survival, cell-to-cell adhesion or cell spreading; therefore, glial tumors with high $^{18}F$-FDG uptake appear to have more aggressive biological properties than those with low uptake.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.15 no.3
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• pp.199-210
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• 1993

Many surgeons are on the point of bone excision and reconstruction of the bone defects by autograft. xenograft, and allograft in the treatment of begin and malignant tumors of bone. Of all type of bone grafts, we received the autograft as the best ideal bone graft. Of autogenic bone graft, replantation of excised autogenic bone for reconstructiong the bone defects has been the ideal method until now, but early bone healing reponses and tumor cell devitalization after replantation of excised autogenic bone have not been identified for clinical applications. So, to evaluate bone healing response after replantation in rabbit's calvarial bone, we divided the experimental group into three groups. Group 1 is a fresh autogenous bone group. Group 2 is a deep frozen group. Group 3 is freeze-dried demineralized group. Obtained result were as followed: 1. Inflammatory cell infiltration appeared at I week and disappeared at 4 weeks in all experimental group, Especially, severe inflammatory cell infiltration showed in fresh autogenous bone group at 2 weeks. Group 3 is the least showing group on the point of inflammatory cell infiltration. 2. Osteoblastic activity evenly increased upto 4 weeks and maintained to 6 weeks and decreased after this period, especially osteoblastic activity in group 2 is less than group 1 and group 3. We can't discriminate between osteoblastic activity of group 1 and that of group 3. 3. In new bone formation, group 3 was more active than any other groups at early stage, but there were little differences among three experimental groups at later state. 4. Bone resorption around the grafted bone slightly appeared at 1 week and disappeared at 4 weeks in all experimental groups. We can find the more bone resorption in group 2 at 2 weeks than any other groups. We could suggest, as appears from our results, that freeze-dried deminiralized bone graft is the useful bone graft in the clinical applications of excised autogenic bone.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.241-248
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• 2014

This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by PMSG and hCG. Two-step EG, DMSO and 4-step EG, DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in EG and DMSO was significantly higher than 4~8 cell (65.4% versus 61.2%, 81.1% versus 72.5%) (p<0.01, p<0.01), but the development rates of 4~8 cell embryos in EG and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4~8 cell embryos in EG were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in EG(77.0% versus 64.4%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The development rate from morular to hatching blastocyst, however, was sinificantly higher in EG than in DMSO during 48 hr (p<0.01). The survival rate of 4~8 cell embryo was 62.5% in EG and 73.3% in DMSO. The development rates of embryo in EG were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. The survival rate of embryo in 2 cell stage was higher than in 4~8 cell stage, and EG appears more effective cryoprotectant than DMSO because EG showed better development rates of embryos in 2 and 4~8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.

• Journal of the Korean GEO-environmental Society
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• v.18 no.12
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• pp.37-45
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• 2017

Structural instability and damage are caused by frost heave during the winter when atmospheric temperature maintains below $0^{\circ}C$. Frost heave is the most representative engineering characteristics of frozen ground and there are various frost susceptibility criteria. Frost susceptibility criteria can be roughly divided into three categories. First, frost susceptibility is determined from particle size distribution, which is practically useful and many countries are adopting. In this paper, several particle size distributions (PSDs) are applied to the frost susceptibility criteria but PSD seems to be not enough to determine whether soils are frost susceptible. Second, it is judged from laboratory frost heave testing results. Laboratory frost heave tests were performed with newly developed thermal controlled triaxial cell and the reliability of frost susceptibility criteria is evaluated. New testing apparatus and method are suitable to meet the existing frost susceptibility criteria. Third, it is compositive frost susceptibility criteria envelope including the particle size distribution, soil classification, and frost heave test. The compositive frost susceptibility criteria envelope should be supplemented based on additional data on various soil types.

• Journal of Embryo Transfer
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• v.18 no.2
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• pp.143-149
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• 2003

This study was carried out to cryopreserve and to investigate characteristics of semen in Hanwoo. Semen was obtained from bulls selected by Daekwanryeong Branch station. Semen was collected each morning of the experiment, placed in water jacketed tubes at 37$^{\circ}C$, and trans-ported to the research laboratory within 10 minutes. Semen was extended with Egg yolk-glycerol extender to contain 50${\times}$10$^{6}$ sperm/ml. Semen was cooled over a 6h period in water jacketed tubes from about 25 to 5$^{\circ}C$, Egg yolk-glycerol extender was added in one step at 5$^{\circ}C$. Semen was aspirated into 0.5ml straws, which were sealed with powder. Egg yolk-glycerol extender, which is used in Hanwoo sperm frozen and stored, semen from 13 Hanwoo bulls collected, the postthawed percentages of motile sperm were 65.7%. In semen characteristics of Hanwoo bulls, number of bulls volume are 5.7 ml and total cell count are 975${\times}$10$^{6}$ m1 ejaculate.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.745-755
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• 2006

The purpose of this study was to investigate the effects of irradiated frozen allogenic bone(IFAB) on the cell proliferation and differentiation of human fetal osteoblasts. Human fetal osteoblasts(hFOB1) were cultured to examine the cellular proliferation for 3 days and 5 days with $1mg/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$, $1{\mu}g/m{\ell}$, $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of IFAB, and to compare the ALP synthesis to control groups for 3 days with DMEM/F-12 1:1 Mixture and $1mg/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$, $1{\mu}g/m{\ell}$, $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of IFAB. To compare the calcium accumulation, hFOBl cultured for 23 days were quantified and photographed. The cellular proliferation of hFOBls treated with IFAB was increased at 5 days to control(p<0.05). The activity of ALP in hFOBls treated with $100ng/m{\ell}$ IFAB was significantly increased at 5 days(p<0.05). A quantified calcium accumulation in hFOBl was significantly increased at $100ng/m{\ell}$, $10ng/m{\ell}$ of IFAB(p<0.05). In the present study, we found that IFAB playa important role of bone formation in the early stage. There was considered that IFAB could be used in the bone graft material.

• Journal of Yeungnam Medical Science
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• v.22 no.2
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• pp.166-182
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• 2005

Background: Cyclin-dependent kinase (CDK) inhibitors are family of molecules that regulate the cell cycle. The CDKN2, a CDK4 inhibitor, also called p16, has been implicated in human tumorigenesis. The CDKN2 inhibits the cyclin/CDK complexes which regulate the transition from G1 to S phase of cell cycle. There is a previous report that homozygous deletion of CDKN2 region on chromosome 9p21 was detected frequently in astrocytoma, glioma and osteosarcoma, less frequently in lung cancer, leukemia and ovarian cancer, but not detected in colon cancer and neuroblastoma. However, little is known about the relationship between CDKN2 and laryngeal cancer. Therefore this study was initiated to investigate the role of CDKN2 in human laryngeal squamous cell carcinoma development.1) Materials and methods: We used 5 human laryngeal carcinoma cell lines whether they have deletions or losses of CDKN2 gene expression by DNA-PCR or RT-PCR, respectively. We examined 8 fresh frozen human laryngeal cancer tissues to detect the loss of heterozygosity (LOH) of CDKN2. PCR was performed by using microsatellite markers of short arm of human chromosome 9 (D9S126, D9S144, D9S156, D9S161, D9S162, D9S166, D9S171, D9S200 and D9SIFNA). For informative cases, allelic loss was scored if the signal of one allele was significantly decreased in tumor DNA when compared to the same allele in normal DNA. Results: The CDKN2 DNA deletion was observed in 3 cell lines. The CDKN2 mRNA expression was observed in only one cell line, which was very weak. LOH was detected in 7 cases (87.5%). Conclusion: These results suggest that CDKN2 plays a role in the carcinogenesis of human laryngeal squamous cell carcinoma.