• Restorative Dentistry and Endodontics
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• v.33 no.4
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• pp.332-340
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• 2008

The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4$^{\circ}C$ for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2. From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.

• Restorative Dentistry and Endodontics
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• v.34 no.4
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• pp.356-363
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• 2009

The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at $4^{\circ}C$for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group. By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.4
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• pp.275-283
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• 2008

Objective: This study was conducted to investigate the effects of the artificial shrinkage and assisted hatching (PZD; patial zona dissetion) before vitrification on the development of vitrified mouse expanding blastocyst. Methods: Mouse 2-cell embryos were collected and cultured in G1.1 and G2.2 to expanding blastocyst. For artificial shrinkage (AS) the micro injection pipette was inserted into blastocoele cavity and blastocoele fluid was aspirated. For assisted hatching (AH) PZD method was used. Control group was -AS/-AH and treatment groups were -AS/+AH, +AS/-AH and +AS/+AH. After AS and AH mouse blastocysts were equillibrated in G10 and G10E20 for 3 mins, respectively, and vitrified in G25E25 after loading on capped pulled-straw. Vitrified mouse blastocysts were thawed and cultured for 24 hrs. The survival and hatching rate was compared among one control and three treatment groups. Results: The survival rates were 99%, 92% in +AS/+AH and +AS/-AH groups and 54%, 58% in -AS/-AH and -AS/+AH group, respectively. The survival rate was significantly higher in +AS group than in -AS group (p<0.01). Hatching rates were 34%, 96% in -AS/-AH and -AS/+AH groups and 41%, 100% in +AS/-AH and +AS/+AH, respectively. The hatching rates was higher in +AH group than in -AH group (p<0.01). After thawing recovery rates were 100%. Loading on capped pulled-straw, that is effective and useful method on vitrification. Conclusion: This study showed that artificial shrinkage of blastocoele cavity and assisted hatching (PZD) significantly improved the development of the vitrified mouse expanding blastocysts.

• Korean Journal of Poultry Science
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• v.41 no.4
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• pp.249-259
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• 2014

Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15% and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05) (85.63% vs 66.81%) by vitrification. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• 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.

• The Korean Journal of Nuclear Medicine Technology
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• v.21 no.1
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• pp.60-64
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• 2017

Purpose Westgard multi-rules application based on test quality improvement and commercialized international standard has been widely used in quality control. However, it is difficult to applicate the Westgard multi-rules in nuclear medicine in vitro tests due to the larger sample sizes and the simultaneous measurement of quality control material and patient sample. This study investigated the usefulness of Westgard multi-rules application in nuclear medicine in vitro tests. Materials and Methods A total of 282 systematic error multi-rules (22s, 101s) recorded in the samsung medical center computer system from January 2013 to June 2016 along with 117 cases of corrective measure record was analyzed. The Quality control implementation is recorded in Hospital information system were divided into 4 high-level areas including quality control material error, experimental procedural error, Kit lot number management error, and others. To prevent quality control material error, the existing method that each staff used their own method was changed. The staff who in charge of managing the quality control material was designated and daily consumption amount of every test was strictly controlled by one person. To prevent other errors, every test step was standardized so that the entire test procedures are identically implemented. Results The total quality control implementation was 117 cases; As a result, 62 quality control material errors were 62 cases, experimental process errors were 24 cases, Kit lot number control errors were 18 cases, and other errors were 13 cases. The quality control material error was corrected and could be used fresh materials within 2 days after thawing. The cases of systemic error were decreased to causes as quality control material error. The quality control materials were reduced above 10 vials to a monthly average. In addition, these errors of experimental processing and Kit lot number were improved by test standardization. Consequently, the cases of 101s and 22s in systematic error rules decreased at least 2 cases to a monthly average. Conclusion To confirm of systematic error through multi-rules application quickly, it is necessary to base on management of the QC material, target values and standard deviation. Moreover, in the event of a systematic error, it was found important to record measures based on test cause analysis. The experiment results are expected to contribute to internal quality control improvement and prompt and accurate result reporting through error recording and causal analysis based on Westgard multi-rules analysis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.8 no.2
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• pp.90-100
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• 1975

The estimation of the pre-processing condition of oyster is of great importance for distributors and processors. This study was attempted to establish the basic data for evaluating the processing suitability of oyster, which is the most important shellfish for domestic use and export. The data were analysed by measuring the condition index, chemical composition and heavy metal content of oysters. In order to eliminate the manual work that has to be done on a tightly closed oyster shell and avoid shrinkage in the oyster meat which is attendant on the steaming process, chemical means to open oyster were examined. finding the method of pretreatment of polyphosphate for frozen oysters were attempted to improve the product quality. The prevention of undesirable color change of the canned oyster meat is another problem to solve. The important results are as follows : 1. The ratio of meat volume and meat weight to the holding capacity by shells may be useful as an index to measure the condition index of oysters. 2. As a whole, monthly changes of moisture and fat content in oysters were reversely correlated. Protein content slightly decreased from April and rapidly decreased in July, and again rapidly increased in August but from September to November decreased slightly. In April, the content of glycogen was 4 percent. From this period to September, glycogen was rapidly decreased. From July to September, it was only 0. 7 to 1 percent but increased from October. There were little seasonal changes in pH value. The pH value of oyster meat was 6.0 to 6.2. The crude ash content was slightly decreased from June to August. 3. The range of monthly change of heavy metal content are as follows: Total mercury was 0 to 0.019 ppm, cadmium was 0.026 to 0.053 ppm, copper was 0.111 to 0.594 ppm, and lead_was 0.061 to 0.581 ppm. 4. By the results of condition index, chemical composition and heavy metal content of oysters, the suitable harvest season as raw materials for processing was the end of December to the end of May of next year. 5. The pretreatment of 10 percent polyphosphate in 5 percent salt solution of oyster meat appeared effective to reduce thawing drip during cold storage. 6. The pretreatment of $Na_2EDTA$ and BHA did not show the color prevention effect to the canned oyster meat during storage. 7. Magnesium chloride affected to open the valves of oysters.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.4
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• pp.602-612
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• 2010

This study was conducted to investigate the state of hygiene education aimed for by bakers, and the evaluations of the importance and the performance concerning sanitary characteristics by the bakers. The questionnaires were administered to 186 bakers in Busan and the data evaluated by 5 scales method of Likert were statistically analyzed. 20.4% of bakers have not received hygiene education. Only 32.8% of bakers have experienced hygiene education regularly. The more the bakers were educated, the more they practiced the contents of hygiene education. The major reason of education unfulfilment was due to insufficient equipments and facilities. The scores of the hygienic performance of educated bakers were significantly (p<0.05) higher than those of uneducated bakers in food sanitation, especially for sanitary characteristics on inspection, pre-preparation and distribution. The mean scores of the importance and the performance evaluated by bakers were 4.05/5.00 and 3.76/5.00, respectively. The bakers assessed the highest scores on the importance and the performance of personal hygiene. The gap score was -0.30 between the importance and the performance for sanitary characteristics. The baker recognized that sanitary management was not performed as much as they recognized its importance. The importance and the performance grid of bakers revealed that the items of checking the certification for the origin of new food ingredient, inspecting deliveries as quickly as possible, separation between preparing and breading time, thawing frozen food under running tap water/in refrigerator, separating disposal gloves and utensils by the purpose, putting products in cleaned and sterilized utensils, letting consumes know the expiration date of products, preventing a rubber hose from being left on the kitchen floor, checking insect nets frequently and so on showed lower scores compared to the mean scores of the importance and the performance. The levels of the hygienic performance by bakers were positively correlated (p<0.01) with the recognitions of the importance on sanitary characteristics. These results might provide basic data for hygienic training and play a role on the improvement of the sanitary management in bakery.

• Korean Journal of Poultry Science
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• v.39 no.4
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• pp.291-295
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• 2012

The use of methylacetamide (MA) as a cryoprotective agent for freezing Korean Native Black rooster Ogye semen was examined with artificial insemination. The diluted Ogye semen with HS-1 was subjected for 2 step dilution method of cryopreservation in which the final concentration of MA was adjusted to 7.5%. The sperm viability after thawing was reduced from $95.17{\pm}0.93%$ to $55.93{\pm}1.38%$ which was confirmed by live-death analysis based on Fluorescence-Activated Cell Sorting (FACS). The rates of fertilized eggs with fresh or frozen-thawed semen were reduced from $94.98{\pm}3.93%$ to $66.36{\pm}8.43%$ at day 7 with significant difference. However, the hatching rates of experiments at day 21 did not shown difference between $92.64{\pm}2.33%$ and $90.45{\pm}8.05%$ (P<0.05). With these results, the utilization of MA for freezing of Ogye spermatozoa could affect on viability of frozen-thawed semen but not on the fertility of lain eggs and hatchability of fertilized eggs and also provide possible tools of freezing for poultry genetic resource conservation.