• 한국식품저장유통학회지
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• 제24권3호
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• pp.343-350
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• 2017

초고압 공정(HPP)은 비가열 공정 중 하나로 식품 중의 세균 증식을 억제하는 방법으로 근래 들어 상업적으로 각광 받고 있다. 자작나무 수액은 여과 및 UV 살균과정을 거쳐 유통되지만 상업적으로 사용하기에는 식품공전 미생물 규격에 적합하지 않아 추가 살균처리 없이 유통을 하기에는 적합하지 않은 단점이 있었다. 본 연구에서는 그 단점을 보완하고 저장유통기한을 연장하기 위해 비가열가공 살균법으로 최근 주목받고 있는 HPP를 적용하고자 하였다. 최적 조건을 설정하기 위해 HPP 조건을 달리 하여 63일 동안 저장하여 변화를 관찰하였고, 그 결과 압력이 높고 처리시간이 길어질수록 미생물제어에 효과적이였다. 설정된 550 MPa, 120 sec의 조건으로 HPP 처리한 후 $5^{\circ}C$와 $10^{\circ}C$에 저장하며 미생물 및 품질변화를 관찰하였다. HPP 미처리군의 경우에는 4-7일차에 급격한 미생물 증식 및 이화학 변화가 일어났지만 HPP 처리군의 경우 $5^{\circ}C$에서 45일, $10^{\circ}C$에서 28일간 저장하는 동안 일반세균, 대장균, 대장균군, 진균 모두 증식이 없어 미생물학적으로 안전한 것으로 나타났다. 또한, pH, 산도, 당도, 탁도 및 갈변도의 변화를 살펴본 결과 $5^{\circ}C$에서는 45일까지 변화가 없었고, $10^{\circ}C$에서는 28일 차에서 pH 감소, 산도, 갈변도, 탁도의 증가로 급격한 변화로 24일 저장하는 것이 이화학적으로도 안정하다고 평가하였다. 이상의 결과로 볼 때 자작나무 수액은 효과적인 살균처리를 하지 않으면 미생물증식, 탁도 및 갈변도 변화, 산도증가로 유통기한이 4일 이하로 저장성이 많이 낮지만, HPP 처리함으로 인해 미생물 증식을 막고 그에 따른 수액의 이화학적 안정성을 확보하여 수액의 품질수명을 기존 미처리군에 비해 6배 연장시킬 수 있을 것으로 판단된다.

• 대한수의학회지
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• 제27권2호
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• pp.331-334
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• 1987

The effects of ethylene glycol as a cryoprotective agent on the survival of frozen-thawed mouse embryos were examined. The effects of the stage of development of ethylene glycol were also examined. Eight-cell embryos, morulae, early blastocysts and mid-blastocysts were recovered from superovulated immature ddY mice. Ethylene glycol was added to the embryos in 5 equal increments 5 minutes apart, giving a final concentration of 1.5M. The embryos were cooled to ${-6^{\circ}C}$ at ${1^{\circ}C/min}$ and seeding was induced at ${-6^{\circ}C}$. After being held for a further 5minutes at the seeding temperature, the samples were cooled to ${-35^{\circ}C}$ at ${0.3^{\circ}C/min}$ and then transferred to liquid nitrogen. Rapid thawing was done by placing the straws in ${37^{\circ}C}$ water. The thawed embryos were diluted in PBS of same time and manners as adding procedures. Survival of 8-cell embryos and morulae were assessed as a normal development of the embryos to the blastocyst stage and expanding blastocyst after 54 hours and 48 hours of in vitro culture, respectively. While those of the early and mid-blastocysts were assessed to the expanding blastocyst stage after 24 hours of in vitro culture. The survival rates of 8-cell embryos, morulae, early blastocysts and mid-blastocysts were 73.8%, 74.3%, 87.8% and 77.4%, respectively. Significant difference on the survival rate among the four stages of development was not observed.

• 한국가축번식학회지
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• 제17권4호
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• pp.271-278
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• 1994

The aim of this study was to compare the two culture systems 1) co-culture with cumulus cells and 2) chemically defined medium supplemented with amino acids (CR1aa) and fetal calf serum (FCS) of in vitro produced bovine embryos from follicular oocytes in vitro. Bovine follicular oocytes were collected from ovaries of slaughtered cows and matured in TCM199 supplemented with 10% FCS and hormones (1$\mu\textrm{g}$/ml FSH-P and 1$\mu\textrm{g}$/ml oestradiol-17$\beta$)24 hours at 39$^{\circ}C$ under 5% CO2 in air. The capacitation of spermatozoa from ejaculated or frozen bull semen was induced by centrifugation through Percoll density gradient (45%, 90%). Then capacitated spermatozoa (1$\times$106/ml) were inseminated into 50${mu}ell$ droplet containing matured follicular oocytes and incubated for 40~42 hours. Cleaved embryos of 2~4cell stage were transferred to the co-culture with cumulus cells and/or CR1aa medium supplemented with FCS. In semen source, the developmental rates to the blastocyst and the hatched blastocyst stages were higher in ejaculated semen(27.6% and 14.9%) than those of frozen-thawed semen(18.3% and 11.8%), respectively. In two culture systems, the proportions of embryonic development upto the blastocysts and the hatched blastocysts were higher of CR1aa medium (22.1% and 12.1%) than those of cumulus cell co-culture (16.8% and 5.1%), respectively. The number of cells in exapnded blastocysts was slightly higher in cumulus cells co-culture (122.6$\pm$8.5) than that in CR1aa medium (117.9$\pm$5.9). The present results indicated that the early development of in vitro produced bovine embryos can be maintained efficiently in CR1aa medium as well as in co-culture with cumulus cells.

• 한국수정란이식학회지
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• 제27권4호
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• pp.259-264
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• 2012

Solution of glycerol, ethylene glycol, sucrose, dextrose (GESD) and cryotop methods were carried out to investigate the survivability on vitrification of embryos. Embryos cultured in vitro were vitrified by GESD of 10 or 8 step and cryotop methods of 6 step, from cryopreservation step to frozen-thawed and culture step. Survival rate and ICM, TE cells of embryos were investigated after frozen-thawed 24 h. As a results, cryotop method was significantly (p<0.05) higher ($85.76{\pm}5.3$ vs. $66.71{\pm}2.4$, $44.80{\pm}2.1%$) than GESD 10 or 8 step methods on survivability. Also, In ICM cell number, cryotop method was significantly (p<0.05) higher to $45.67{\pm}4.7$ cells than GESD 8 step method. TE cell number was significantly (p<0.05) highest to $111.00{\pm}11.0$ cells in cryotop method. On the other hand, survival rate, TE and total cell number were all the significantly (p<0.05) high, except ICM in GESD 10 step method between GESD 10 step method and GESD 8 step method. In conclusion cryotop method was to be most effective, but it is considered necessary to study vitrification method for step-by-step freezing and thawing process.

• Clinical and Experimental Reproductive Medicine
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• 제45권1호
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• pp.25-30
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• 2018

Objective: This study conducted a preliminary examination of the effects of three-area laser-assisted zona thinning (LAZT) during the cleavage stage of embryo development on the hatching process in human in vitro fertilization-embryo transfer (IVF-ET) with subjects of advanced female age or frozen-thawed (FT) embryos. Methods: Eight-cell stage embryos were treated with LAZT in three areas of the zona pellucida at $120^{\circ}$ intervals. The control group was embryos without LAZT. Of the 72 consecutive fresh cycles and the 28 FT embryo transfer cycles, the patients in 55 fresh cycles and 17 FT cycles declined LAZT, and those cycles were defined as the control group. Results: In the fresh cycles, the pregnancy rates were similar in the LAZT and control groups. However, in the FT cycles, the pregnancy rate was significantly higher in the LAZT group than in the control group (45.5% in the LAZT group vs. 23.5% in the control group, p< 0.05). Conclusion: These results show that multi-area LAZT resulted in significantly improved pregnancy outcomes in human 8-cell embryos compared to controls.

• 한국수정란이식학회지
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• 제18권3호
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• pp.179-186
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• 2003

Human embryonic stem (hES) cell lines have been derived from human blastocysts and are expected to have far-reaching applications in regenerative medicine. The objective of this study is to improve freezing method with less cryo-injuries and best survival rates in hES cells by comparing various vitrification conditions. For the vitrifications, ES cells are exposed to the 4 different cryoprotectants, ethylene glycol (EG), 1,2-propanediol (PROH), EG with dime-thylsulfoxide (DMSO) and EG with PROH. We compared to types of vehicles, such as open pulled straw (OPS) or electron microscopic cooper grids (EM grids). Thawed hES cells were dipped into sequentially holding media with 0.2 M sucrose for 1 min, 0.1 M sucrose for 5 min and holding media for 5 min twice and plated onto a fresh feeder layer. Survival rates of vitrified hES cells were assessed by counting of undifferentiated colonies. It shows high survival rates of hES cells frozen with EG and DMSO (60.8％), or EG and PROH(65.8％) on EM grids better than those of OPS, compared to those frozen with EG alone (2.4％) or PROH alone (0％) alone. The hES cells vitrified with EM grid showed relatively constant colony forming efficiency and survival rates, compared to those of unverified hES cells. The vitrified hES cells retained the normal morphology, alkaline phosphates activity, and the expression of SSEA-3 and 4. Through RT-PCR analysis showed Oct-4 gene expression was down-regulated and embryonic germ layer markers were up-regulated in the vitrified hES cells during spontaneous differentiation. These results show that vitrification method by using EM grid supplemented with EG and PROH in hES cells may be most efficient at present to minimize cyto-toxicity and cellular damage derived by ice crystal formation and furthermore may be employed for clinical application.

• Journal of Microbiology and Biotechnology
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• 제6권2호
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• pp.128-131
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• 1996

Lactobacillus acidophilus KFRI 233 (of human origin) exhibited a high tolerance to bile. The maximum cell yield was 6.6${\time}10^9 CFU$ per gram of whey in a 5.0% whey medium. Cell growth was improved with the addition of 0.5% thiotone and 0.25% calcium carbonate. Cell growth reached a maximum level of 5.4${\times}10^8$ CFU/ml at 20 h. Eighty-nine percent of the viable cells in the centrifuged concentrate survived freezing at $-70^{\circ}C$ and this frozen concentrate showed no reduction in the viable cell count after 30 days at $-70^{\circ}C$. Eight percent of the viable cells survived freeze-drying after the addition of 1 g/l sodium carbonate before harvesting by centrifuging and this freeze-dried concentrate showed only a slight reduction in the viable cell count after 30 days at $4^{\circ}C$.

• 대한두경부종양학회지
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• 제36권2호
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• pp.41-44
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• 2020

Clear cell hidradenoma is a skin adnexal tumor originating from eccrine glands. The risk of local recurrence after surgical resection exceeds 50%, and 6-19% of cases are malignant. The rarity of clear cell hidradenoma and its diverse histological findings make this type of tumor a diagnostic challenge. We present a case of recurrent clear cell hidradenoma of the posterior neck in a 70-year-old woman. The tumor recurred once after complete excision, and did not recur again after 1-cm wide excision and reconstruction with a local bilobed flap. Recurrent clear cell hidradenomas are activated by surgical stimulation, increasing the risk for metastasis. Therefore, we suggest that wide excision with confirmation of a tumor-free margin by frozen-section biopsy should be the first-line treatment for recurrent benign clear cell hidradenoma.

• 대한세포병리학회지
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• 제17권1호
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• pp.32-37
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• 2006

The objective of this study was to evaluate the cytomorphologic features of histologically confirmed clear cell carcinoma of the ovary and to evaluate the applicability of scrape or fine-needle aspiration cytology in making an intraoperative diagnosis. We reviewed scrape or fine-needle aspiration cytology findings in tissues taken from 6 patients with clear cell carcinoma of the ovary. The cytologic diagnosis was based primarily on findings in alcohol-fixed, hematoxylin-eosin (H-E) stained smears. The formation of material resembling a basement membrane was a characteristic finding in these smears. This extracellular hyaline material was stained light pink with H-E and was frequently found within tumor cell clusters as well as in the background material. Multinucleated giant cells were found occasionally. Each tumor cell had an abundant, clear, or granular cytoplasm with a distinct cellular membrane. Scrape cytology is a simple and rapid supportive method and could be helpful in diagnosing clear cell carcinoma of the ovary, especially when marked artifacts appear in the frozen section.

• 대한두경부종양학회지
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• 제13권2호
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• pp.206-212
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• 1997

H$\"{u}$rthle cell neoplasm of the thyroid gland is an uncommon, but potentially malignant lesion. However, in many instances, the malignant potential of the H$\"{u}$rthle cell neoplasm is very difficult to judge histologically. For this reason, the biologic behavior of this tumor and its optimal treatment have come under considerable debate in recent years. In order to review the clinicopathologic features of the H$\"{u}$rthle cell neoplasm and to determine its optimal treatment modalities, we studied 26 patients with path logical proof of H$\"{u}$rthle cell tumor from January 1987 to September 1997. We also performed an immunohistochemical study using the monoclonal antibodies against antigen CD34 for the angiogenic activity of this tumor and evaluated the differences of microvessel density(MVD) between benign and malignant tumors. The age of the patients ranged from 1 to 71 years with a mean of 44.2 years. There were 6 males and 20 females(M : F= 1 : 3.3). The accuracies of fine needle aspiration biopsy and frozen section were very low; 6.3% and 34.8%, respectively. There were 20 benign tumors and 6 malignant tumors(23.1%). All the malignant tumors were microinvasive(intermediate) type which had minimal capsular invasion and most of them(5 cases) were diagnosed postoperatively. Any specific clinicopathologic differences were not seen between benign and intermediate groups. Most of the cases had conservative surgeries(15 ipsilateral lobectomy-isthmusectomy, 7 subtotal thyroidectomy) while total thyroidectomy was performed in 4 cases. Of the cases with malignant tumor, 2 had ipsilateral lobectomy-isthmusectomy, 3 had subtotal thyroidectomy and the remaining 1 had total thyroidectomy. Mean size of the tumors was 3.0 cm(0.1- 8.5 cm) in the greatest diameter and multiple tumors were seen in 6 cases(23.1 %). During the follow-up period, only one recurrence(3.8%) of benign tumor occurred but distant metastasis or cause-specific death was seen in the benign or intermediate groups. Mean MVDs of the benign(n=13) and intermediate(n=6) groups were $121.7{\pm}35.3$ and $114.3{\pm}31.7$, respectively and there was no statistical significance between them. In conclusion, because of the low accuracies of fine needle aspiration biopsy and frozen section for the H$\"{u}$rthle cell neoplasm, the extent of surgery could be individualized based on permanent pathologic examination; Conservative surgery would be adequate for patients with benign or intermediate H$\"{u}$rthle cell neoplasm and total or near-total thyroidectomy for those with definite malignancy.