• Title/Summary/Keyword: frozen cell

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Effect of the Culture Media of Bovine Oviductal Epithelium on Development of the Early Bovine Embryos Derived from in vitro Fertilization (소 난관상피세포배양액이 체외수정 유래 분할란의 발육에 미치는 영향)

  • Park Jong-Im;Hwang Woo-Suk;Jo Choong-Ho;Lee Byeong-Chun
    • Journal of Veterinary Clinics
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    • v.9 no.1
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    • pp.323-332
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    • 1992
  • The present study was carried out to examine the effect of oviduct epithelium and its conditioned medium on e development of early bovine embryos in vitro. Oocytes obtained from ovarian follicles of slaughtered cows were cultured in TCM199 with 10% fetal calf serum for 22-24hrs and then fertillzed in vitro using frozen-thawed semen treated with BO-caffein, BO-BSA(20mM heparin added). Oviduct epithelium was collected in each stage of the estrus cycle and conditioned medium was the medium in which oviduct epithelium in early luteal stage was cultured. In vitro fertilized bovine embryos of 1~2 cell were co-cultured with oviduct epithelium from different estrus cycles, cultured in conditioned medium, and cultured in rabbit oviduct. The cleavage rates of in vitro fertilized early bovine embryos co-cultured with oviduct epithelial cell from early luteal, luteal and follicular phase of estrus cycle(67.2~70.8%) and cultured in conditioned medium(56.7%) were significantly(p<0.05) higher than that of the control(44.2%) The rate of development to morula or blastocyst stage in oviduct epithelial cell co-culture(15.3~32.5%) from three phase of estrus cycles and conditioned medium(14.5%) were significantly(p<0.05) higher than that of the control(5.2%). The oviduct epithelial cell from early luteal phase gave a significantly( p<0.05) higher rate of development to morula or blastocyst stage than both luteal and follicular phase. The results of in vivo culture in rabbit oviduct of early bovine embryos were 52.1% for the cleavage rate and 26.7% for the rate of development to morula or blastocyst stage.

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ANTI-TUMOR AND ANTI-ANGIOGENIC EFFECT OF THALIDOMIDE ON ORAL SQUAMOUS CELL CARCINOMA XENOGRAFTS IN NUDE MICE (누드마우스에 이종이식된 구강편평상피세포암종에 대한 thalidomide의 항암효과와 혈관형성억제에 관한 연구)

  • Kim, Su-Gon;Myoung, Hoon;Kim, Myung-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.4
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    • pp.330-336
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    • 2001
  • Angiogenesis is an essential process for the growth, invasion and metastasis of cancer. However, it is uncertain that antiangiogenic effects can be a major treatment strategy of oral cancer. The aim of this study was to investigate whether thalidomide, which is known to be a potent inhibitor of angiogenesis, have inhibitory effect on the growth and antiangiogenic effects of oral squamous cell carcinoma(OSCC) xenografted in nude mice and whether antiangiogenesis of thalidomide can be included as a major treatment strategy of oral cancer. After human oral squamous cell carcinoma strain KB was subcutaneously implanted in 20 nude mice, the volume of tumor was measured every three days. When the tumor mass reached $75{\sim}100mm^3$, thalidomide(200mg/kg/d) was administered into 10 experimental nude mice and the same volume of distilled water was administered into 10 control nude mice and the tumor volume was measured every three days. The excised tumor masses on the 30th day after administration were frozen and processed for immunohistochemistry using vascular endothelial growth factor(VEGF) and CD31. We evaluated microvessel density and VEGF expression. The results were as follows ; 1. Thalidomide retarded the growth of human OSCC as compared with the control group, but it was not statistically significant. 2. A statistically significant lower microvessel density was observed in the thalidomide-treated group than in the control group(p<0.01) and thalidomide significantly reduced VEGF expression (p<0.01). Thalidomide exhibited significantly antiangiogenic effect, but did not inhibit the growth of human OSCC effectively. Antiangiogenic therapy of thalidomide alone is not likely to be effective in the treatment of human OSCC, but might be regarded as adjuvant chemotherapeutic strategy.

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Involvement of CELSR3 Hypermethylation in Primary Oral Squamous Cell Carcinoma

  • Khor, Goot Heah;Froemming, Gabrielle Ruth Anisah;Zain, Rosnah Binti;Abraham, Thomas Mannil;Lin, Thong Kwai
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.219-223
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    • 2016
  • Background: Promoter hypermethylation is a frequent epigenetic mechanism for gene transcription repression in cancer and is one of the hallmarks of the disease. Cadherin EGF LAG seven-pass G-type receptor 3 (CELSR3) contributes to cell contact-mediated communication. Dysregulation of promoter methylation has been reported in various cancers. Objectives: The objectives of this study were to investigate the CELSR3 hypermethylation level in oral squamous cell carcinomas (OSCCs) using methylation-sensitive high-resolution melting analysis (MS-HRM) and to correlate CELSR3 methylation with patient demographic and clinicopathological parameters. Materials and Methods: Frozen tissue samples of healthy subjects' normal mucosa and OSCCs were examined with regard to their methylation levels of the CELSR3 gene using MS-HRM. Results: MS-HRM analysis revealed a high methylation level of CELSR3 in 86% of OSCC cases. Significant correlations were found between CELSR3 quantitative methylation levels with patient ethnicity (P=0.005), age (P=0.024) and pathological stages (P=0.004). A moderate positive correlation between CELSR3 and patient age was also evident (R=0.444, P=0.001). Conclusions: CELSR3 promoter hypermethylation may be an important mechanism involved in oral carcinogenesis. It may thus be used as a biomarker in OSCC prognostication.

Cryopreservation of rabbit embryos by vitrification (Vitrification 방법에 의한 토끼수정란의 동결에 관한 연구)

  • Choe, Sang-yong;Lee, Young-rak;Rho, Gyu-jin;Lee, Hyo-jong;Park, Choong-saeng
    • Korean Journal of Veterinary Research
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    • v.35 no.3
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    • pp.635-641
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    • 1995
  • The purpose of this study was to investigate the effects of developmental stage and equilibration time on survival of rabbit embryos following freezing by vitrification. Adult New Zealand White female rabbits were superovulated with PMSG and hCG. The 8-cell stage embryos were collected from 40 to 45 hours after hCG injection by flushing oviducts with Dulbecco's phosphated buffered saline and in vitro cultured in TCM-199 containing 10% fetal calf serum(FCS). Each embryos developed in vitro to 16-cell, compact morula and blastocyst was cryopreserved and cultured following thawing to examine their developmental potential to expanded blastocyst stage in vitro. The frozen-thawed-cultured embryos were stained with Hoechst 33342, and their nuclei were counted using a fluorescence microscope. On the toxicity test of EFS solution as cryopreservation, the survival rates of 8-cell stage embryos was decreased in reverse to increasing of exposure time over 5 minutes. The post-thaw survival rates of embryos on equilibration times was significantly(P<0.05) higher for 2 min. than for 5 or 10 minutes. From morula to blastocyst of rabbit embryos was more suitable than 8-cell stage for cryopreservation by vitrification. The higher post-thaw survival rate of embryos can be achieved by keeping the cryoprotectant at $4^{\circ}C$ than at $20^{\circ}C$. The mean number of nuclei per embryo following freezing by vitrification and in vitro culture to expanded blastocyst at compacted morula and blastcyst was not significantly differ from fresh blastocyst.

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Rapid freezing versus Cryotop vitrification of mouse two-cell embryos

  • Inna, Namfon;Sanmee, Usanee;Saeng-anan, Ubol;Piromlertamorn, Waraporn;Vutyavanich, Teraporn
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.3
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    • pp.110-115
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    • 2018
  • Objective: To compare our in-house method of embryo freezing with Cryotop vitrification in terms of immediate survival, subsequent cleavage and blastocyst formation, and cell numbers in blastocysts. Methods: Two-cell mouse embryos were randomly allocated into three groups: a non-frozen control group (group 1, n = 300), a group that underwent Cryotop vitrification (group 2, n = 300), and a group that underwent our in-house freezing method (group 3, n = 300). Results: There were no significant differences between groups 2 and 3 in the immediate survival rate (96.3% vs. 98.6%, respectively; p= 0.085), the further cleavage rate (91.7% vs. 95.0%, respectively; p= 0.099), or the blastocyst formation rate (80.7% vs. 78.6%, respectively; p= 0.437). The cell numbers in the blastocysts from groups 1, 2, and 3 were comparable ($88.99{\pm}10.44$, $88.29{\pm}14.79$, and $86.42{\pm}15.23$, respectively; p= 0.228). However, the percentage of good-quality blastocysts in the Cryotop vitrification group was significantly higher than in the group in which our in-house method was performed, but was lower than in the control group (58.0%, 37.0%, and 82.7%, respectively; p< 0.001). Conclusion: At present, our method is inferior to the commercial Cryotop vitrification system. However, with further improvements, it has the potential to be useful in routine practice, as it is easier to perform than the current vitrification system.

Malaria Cell Image Recognition Based On VGG19 Using Transfer Learning (전이 학습을 이용한 VGG19 기반 말라리아셀 이미지 인식)

  • Peng, Xiangshen;Kim, Kangchul
    • The Journal of the Korea institute of electronic communication sciences
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    • v.17 no.3
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    • pp.483-490
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    • 2022
  • Malaria is a disease caused by a parasite and it is prevalent in all over the world. The usual method used to recognize malaria cells is a thick and thin blood smears examination methods, but this method requires a lot of manual calculation, so the efficiency and accuracy are very low as well as the lack of pathologists in impoverished country has led to high malaria mortality rates. In this paper, a malaria cell image recognition model using transfer learning is proposed, which consists in the feature extractor, the residual structure and the fully connected layers. When the pre-training parameters of the VGG-19 model are imported to the proposed model, the parameters of some convolutional layers model are frozen and the fine-tuning method is used to fit the data for the model. Also we implement another malaria cell recognition model without residual structure to compare with the proposed model. The simulation results shows that the model using the residual structure gets better performance than the other model without residual structure and the proposed model has the best accuracy of 97.33% compared to other recent papers.

Cryopreservation of Porcine Embryos using Glycerol, Egg Yolk and Trehalose (Glycerol, 난황 및 Trehalose를 이용한 돼지유정란의 동결)

  • 장원경;박수봉;이명식;김태현;박용윤;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.43-48
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    • 1995
  • The purpose of this study was to examine the suvival rates of cryopreserved porcine embryos collected on Day 6 (Day 0=onset of estrus) with various cryprotectants. Eighty two embryos at different stages, ranged from expanded blastocyst to hatched blastocyst, were allocated to 6 experimental groups in different combinations of cryoprotectants glycerol, egg yolk and/or trehalose. Porcine embryos were cryopreserved using conventinal slow freezing precedures. The embryos were equilibrated with one of the freezing solutions, cooled from 25 to -7$^{\circ}C$ at 1$^{\circ}C$/min, seeded at -7$^{\circ}C$ frozen to -36$^{\circ}C$ at 0.5$^{\circ}C$/min, and then plunged into liquid nitrogen. The frozen embryos were thawed by immersion in 37$^{\circ}C$ water and the cryoprotectants were removed by dilution with 0.5M sucrose solution. Embryonic survival was estimated from the normal development of embryos for 12, 24 and 48hrs culture. Then the embryos were stained and their cell nuclei was counted. The survival rates of morphological embryos were significantly higher in group I (10% glycerol) or group IV (10% glycerol+10% egg yolk+0.5M trehalose) than those in other groups, although the nuclei number was quite higher in embryos treated with 10% glycerol and 0.25M trehalose at expended blastocyst. However, hatched blastocysts showed higher viability and nuclei number treated with either egg york or trehalose, but the survival rates after 48hrs of culture were quite low. These results indicate that egg yolk and trehalos as a supplement to freezing solutions can be useful to the cryopreservation ofn porcine embryos.

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Macrofollicular Variant of Papillary Thyroid Carcinoma with Extensive Hemorrhage -Report of A Case- (출혈을 동반한 대여포성 유두상 갑상선 암종 -1예 보고-)

  • Kim, Hae-Ryoung;Lee, Kwang-Gil;Kim, Eun-Kyung;Park, Cheong-Soo;Chung, Woung-Youn;Yang, Woo-Ick;Hong, Soon-Wong
    • The Korean Journal of Cytopathology
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    • v.15 no.1
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    • pp.60-64
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    • 2004
  • The macrofollicular variant of papillary thyroid carcinoma (MVPC) is characterized by macrofollicles occupying more than half of the tumor and demonstrating nuclear features of classic papillary carcinoma. It is difficult to recognize on fine needle aspiration (FNA) cytology due to the paucity of aspirated neoplastic cell clusters, especially when the tumor is associated with extensive areas of hemorrhage. Case: A 34-year-old female presented with a well-demarcated nodule in the thyroid gland, diagnosed as a benign nodule on ultrasonography and computed tomography. FNA cytology smear revealed a few small aggregates of follicular cells with morphological features suspicious for papillary carcinoma, set in a background of hemorrhage, inflammatory cells, and hemosiderin-laden macrophages. Intraoperative frozen section revealed macrofollicular nests filled with hemorrhage and composed of follicular cells demonstrating nuclear clearing and grooves. Conclusion: MVPC is a rare but distinctive variant of papillary carcinoma, which is easily mistaken for adenomatous goiter or benign macrofollicular neoplasm on radiologic findings. The cytopathologist should alert oneself on encountering benign radiologic findings and any smear composed of scant numbers of follicular cells with nuclear features suspicious for papillary carcinoma despite the bland-looking background of hemorrhage and hemosiderin-laden macrophages, and recommend intraoperative frozen sections for a definite diagnosis.

Effects of cryoprotectants and sucrose concentrations on the viability of aggregated mouse embryos frozen rapidly in liquid nitrogen vapour (동결보호제 및 Sucrose 농도가 급속동결한 마우스 집합배의 생존성에 미치는 영향)

  • Shin, Sang-tae
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.523-527
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    • 1991
  • The effects of ethylene glycol, DMSO and glycerol as cryoprotectants and the effect of concentrations(0, 0.25, 0.5 and 1.0M) of sucrose in the diluent on the viability of the aggregated morulae frozen rapidly in liquid nitrogen$(LN_2)$ vapour were examined. The morulae were produced by aggregation of ICR and CBA mice embryos at 8-cell stage. Before freezing the embryos were equilibrated in 1.5M cryoprotectants+0.25M sucrose in oae-step or in 3.0M cryoprotectants+0.25M sucrose in two-steps. The embryos were pipetted into the freezing medium fraction of 0.25ml plastic straws. The straws were frozeu by directly transfer into $LN_2$ vapour(about lcm above $LN_2$) for 2 minutes, and then plunged into $LN_2$. After thawing the cryoprotectants were diluted with 0, 0.25, 0.5 or 1.0M sucrose solution. The post-thawed in vitro viability of the aggregated embryos was significantly dependent on the type and concentration of cryoprotectants in the freezing medium and also on the concentration of sucrose in the diluent. When the aggregated embryos were equilibrated in 1.5M cryoprotectants +0.25M sucrose in one-step and diluted with 0.5M sucrose after thawing, the survival rate of the embryo5 was significantly(p<0.05) higher in DMSO(62.5%) or ethylene glycol(52.2%) than in glycerol(33.3 %). In the case that the concentration of the cryoprotectants was raised to 3.0M in two-steps, thc higher survival rate of the embryos was obtained in ethylene glycol or glycerol than in DMSO followed by diluting them with 0.5 or 1.0M sucrose after thawing(p<0.01).

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Effects of Rosemary Extract, ${\alpha}-Tocopherol$ and Vacuum Packaging on Qualities of Herring Fillet during Cold and Frozen Storage (Rosemary 추출물, ${\alpha}-Tocopherol$ 및 진공 포장이 청어 Fillet의 냉장 및 동결 저장 중 품질에 미치는 효과)

  • Yang, Seung-Taek;Park, Sang-Woo
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.697-704
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    • 1999
  • To investigate the combination effects of natural preservatives such as rosemary extract and ${\alpha}-tocopherol$, and vacuum packaging method on the shelf-life of herring (Clupea pallasi) fillet during cold and frozen storage, quality attributes including moisture content, pH, K-value, volatile basic nitrogen (VBN), viable cell count, peroxide value (POV) and color value were analyzed. Good qualities of the samples with air and vacuum packaging stored at $4^{\circ}C$ were maintained at least for 3 days (control; one day in air) and for 7 days (control; 3 days in vacuum), and those of the samples stored at $-20^{\circ}C$ were for 90 days (control; 80 days in air, 90 days in vacuum), while those of the samples stored at $20^{\circ}C$ deteriorated in one day.

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