• Title/Summary/Keyword: thawing conditions

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Effect of Equilibration Time and Cell Stage on the Survival of IVF Bovine Embryos Cryopreserved by Vitrification (한우 체외수정란의 동결보존시 평형시간과 배 발달단계가 생존성에 미치는 영향)

  • 공일근;주영국;이은봉;김용권;박충생
    • Journal of Embryo Transfer
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    • v.9 no.1
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    • pp.7-14
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    • 1994
  • The present experiments on cryopreservation were designed to examine the effects of solution toxicity, equilibration time and cell stages on the post-thaw survival of bovine IVF embryos. The oocytes were matured in vitro(IVM) for 24 hrs. in TCM-199 supplemented with 35 $\mu$g /ml FSH, 10 $\mu$g /ml LH, 1 $\mu$g /ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro(IVC) with bovine oviductal epithelial cells for 7 to 9 days. The bovine IVF embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen, and thawed rapidly. 1. after the bovine blastocysts were exposed to EFS solution for 2 min. at room temperature and then they were washed in 0.5 M sucrose solution and TCM-199, they were cultured to examined cryoprotectant induced injury during exposure, Most of the embryos(95.0%) developed to reexpanded blastocoels. However, when the exposure time was extended to 5 and 10 min, these development rates dropped dramatically in 5 min. (69.5%) and 10 min. (47.4%), respectively, 2. When the bovine IVF embryos were vitrified in EFS solution after the equilibration for 1 and 2 min. exposure, The embryos to have reexpanded blastocoels following thawing, washing and culture processes were found to he 82.6 and 73.9%, respectively. However, when the exposure time was extended to 3 min, this survival rate dropped to 18.2%. The optimal time for equilibration of bovine IVF blastocysts in EFS solution seemed to he 1~2 min. 3. When the bovine IVF embryos were equilibrated for 1 min. the significantly (P<0. 05) higher post-thaw survival rates were obtained from the embryos of blastocyst stage(81.3%) than morulae stage(5. 1%). The optimal cell stage for viterification with EFS solution proven to he blastocyst stage in bovine IVF embryos. 4. The number of blastomeres of blastocyst stage was examined with nuclear staining with Hoechst 33342 during 7 to 9 days post-insemination. The cell counts of frozen bovine IVF embryos were found significantly(P$\geq$7.5 and those of the fresh embryos 76.6$\geq$7. 1, which were cultured in the sarne period and conditions as frozen embryos.

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Study on Adhesive Strength of Polymer Modified Cement Mortar for Maintenance in Concrete Structure (콘크리트 구조물 보수용 폴리머시멘트 모르타르의 부착강도 특성에 관한 연구)

  • Park, Sang-Soon;Kim, Jung-Heum
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.14 no.5
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    • pp.128-135
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    • 2010
  • Polymer-modified cement mortar(PCM) has been widely used for strengthening of the concrete structures due to its excellent physical properties such as high strength and durability. Adhesive strength or behavior, on the other hands, between PCM and concrete is very important in strengthening the concrete member using PCM. Therefore the adhesive failure mechanism between PCM and concrete should be fully verified and understood. This study was performed to evaluate adhesive strength of PCM to the concrete by the direct pull-out test. In the direct pull-out tests, the adhesive strength under the various pre-treatment conditions such as immersion, thunder shower, freezing and thawing are evaluated. Also, the field direct pull-out test are performed to investigate the adhesive strength of mock-up test specimens. In the results of the test, the adhesive strength value by field test are lower than those of the standard curing condition. From these comparison and investigation, field test result was similar with the thunder shower test result. The results of the test was used to evaluate the korean industrial standard of polymer modified cement mortars for maintenance in concrete.

Evaluation of Durability Performance in Concrete Incorporating Low Fineness of GGBFS (3000 Grade) (저분말도 고로슬래그 미분말(3000급)을 혼입한 콘크리트의 내구성능평가)

  • Lee, Seung-Heun;Cho, Sung-Jun;Kwon, Seung-Jun
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.23 no.4
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    • pp.96-102
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    • 2019
  • When GGBFS(Ground Granulated Blast Furnace Slag) with high blaine is incorporated in concrete, compressive strength in the initial period is improved, but several engineering problems arise such as heat of hydration and quality control. In this paper, compressive strength and durability performance of concrete with 3,000 Grade-low fineness slag are evaluated. Three conditions of concrete mixtures are considered considering workability, and the related durability tests are performed. Although the strength of concrete with 3,000 Grade slag is slightly lower than the OPC(Ordinary Portland Cement) concrete at the age of 28 days, but insignificant difference is observed in long-term compressive strength due to latent hydration activity. The durability performances in concrete with low fineness slag show that the resistances to carbonation and freezing/thawing action are slightly higher than those of concrete with high fineness slag, since reduced unit water content is considered in 3,000 Grade slag mixture. For the long-term age, the chloride diffusion coefficient of the 3000-grade slag mixture is reduced to 20% compared to the OPC mixture, and the excellent chloride resistance are evaluated. Compared with concrete with OPC and high fineness GGBFS, concrete with lower fineness GGBFS can keep reasonable workability and durability performance with reduced water content.

Effect of Maximum Aggregate, Porosity, and Temperature on Crack Resistance and Moisture Susceptibility of Porous Asphalt Mixtures (최대입경, 공극률, 온도가 다공성 아스팔트 혼합물의 균열저항성 및 수분민감성에 미치는 영향)

  • Yoo, In-Kyoon;Lee, Su-Hyung;Park, Ki-Soo;Yoon, Kang-Hoon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.3
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    • pp.611-619
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    • 2021
  • Porous asphalt pavement (PAP) has many functions, such as reducing accidents and decreasing noise. On the other hand, vulnerability is inevitable because PAP contains approximately 20% porosity. This study evaluated the effects of the maximum aggregate size (MAS), temperature, and porosity on the PAP durability. The indirect tensile strength measures durability. This study tested the samples that stayed dry and were moisturized by freezing and thawing for mixtures having the same porosity of 20% and MAS of 13mm, 10mm, and 8mm. The same test was performed on a mixture of 20% and 22% voids made of the same material with a MAS of 10mm. As a result, for 20% porosity, significant differences in the changes in MAS and temperature were found. A clear difference was observed between 8mm and 13mm under dry conditions, but there were no other significant differences in the MAS change. Furthermore, there was a clear difference in temperature for the change in porosity and temperature, but the gap in 2% porosity at 20% did not show a clear difference. Therefore, it is necessary to develop a more durable PAP through quantitative evaluations of the factors affecting the PAP durability.

A simple mid-term preservation method (SMPM) of plant callus under low temperature conditions (저온 보존을 이용한 간편 중기 식물캘러스 저장법)

  • Park, Sung-Chul;Park, Su Hyun;Kim, Soyoung;Jeong, Yu Jeong;Kim, Cha Young;Jeong, Jae Cheol
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.187-192
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    • 2022
  • The repeated monthly or weekly subculture of plant callus is labor intensive and increases the risk of somaclonal variation from the parental callus line. The most effective method for preserving plant callus is cryopreservation, which involves storage in liquid nitrogen. However, this method cannot be applied to the callus of different plant species in the same manner, so it is difficult to develop a standardized cryopreservation method. In addition, the survival rate of the frozen callus after thawing and the regeneration rate after survival are uncertain. Therefore, it is necessary to develop a method to extend the subculture interval of plant callus in an active state. In this study, active plant calli of various species without freezing was incubated at 15℃ for 4 to 12 weeks without subculture. After 12 weeks, 8 lines of plant callus grew less than 2-fold when cultured at 25℃, but at least 2 times as much when cultured at 15℃. Moreover, total antioxidant activity did not differ significantly between plant callus recovered at 25℃ after culturing at 15℃ or at 25℃. These results show that the subculture interval can be extended at a temperature of 15℃ without need for modified medium composition or additional processes. In addition, positive results in all calli of several plant species are expected to reduce labor as well as somaclonal variation by increasing the subculture.

Biogeochemistry of Methane in Water and Sediment: Methane Generation in Coastal Areas with Bottom Water Hypoxia (메탄의 생지화학적 거동과 한국 연안해역 저(빈)산소 층 발달에 따른 메탄 생성)

  • DONGJOO JOUNG
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.28 no.3
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    • pp.95-120
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    • 2023
  • Methane (CH4) is a key greenhouse gas in the atmosphere with 85 times greater greenhouse potent relative to carbon dioxide (CO2). The atmospheric concentration of CH4 is rapidly increasing due to the intensive usage of CH4 and the thawing of the cryosphere. Additionally, with the current warming of ocean water, the dissociation of gas hydrates, an ice-like compound and the largest reservoir of CH4 on Earth, is expected to occur, resulting in the release of CH4 from the seafloor into the overlying water and atmosphere. Moreover, bottom water hypoxia is another concern that potentially introduces greenhouse gases into the atmosphere. With ongoing global warming and eutrophication, the size and duration of bottom water hypoxia are rapidly increasing. These low-oxygen conditions would relocate the redox zone shallower in sediment or in the water column, causing the release of CH4 into the atmosphere and thereby intensifying global warming. However, there exists a gap in the understanding of CH4 dynamics including its generation in relation to bottom water hypoxia. Therefore, this review article aims to understand the relationship between CH4 and bottom water hypoxia and to draw attention to CH4 investigation in Korea.

Current Status of Sanitation Management Performance in Korean-Food Restaurants and Development of the Sanitary Training Posters Based on their Risk Factors (한식당의 위생관리 현황 평가 및 위험요인 중심의 위생교육용 포스터 개발)

  • Kim, Sun-Jung;Yi, Na-Young;Chang, Hye-Ja;Kwak, Tong-Kyung
    • Journal of the Korean Society of Food Culture
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    • v.23 no.5
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    • pp.582-594
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    • 2008
  • This study aimed at evaluating current sanitation management performances in Korean-Food restaurants by their operation types and to develop sanitary training posters based on the risk factors, in an attempt to improve the level of sanitation management in Korean food service facilities. Eighteen Korean-food restaurants that are managed by franchisor, franchisees as well as self-managed with large-scale and small-scale restaurants in Seoul and Gyeonggi-Do, were evaluated by on-the-spot inspectors with an auditing tool consisting of three dimensions, nine categories and thirty four items. Data were analyzed using SPSS. The total score of each group showed that restaurants managed by franchisees ranked the highest (59 out of 100 points), while self-managed, small-scale restaurants ranked the lowest (44 out of 100 points). In the categorization of sanitation management compliance, the dimensions of food hygiene during production recorded the lowest compliance rate of 47.7% (22.89/48.0 points) followed by the dimension of environmental hygiene 59.3% (20.17/34.0 points) and personal hygiene 60.5% (10.89/18.0 points). This indicated the need for urgent improvement. The items which showed the lowest compliance rates were 'proper thawing of frozen foods' (0%), 'notifying and observing heating/reheating temperature' (6%), 'using of hand-washing facility and proper hand-washing' (33%), 'monitoring temperature of frozen-foods and cold-foods' (35%), and 'prevention of cross-contamination' (36%) among thirty four items. Self-managed, small-scale restaurants, in particular, needed to improve sanitary practices such as 'sanitation education for employee', 'verifying the employee health inspection reports', 'storing food on the shelves 15 cm distance away from the wall', 'suitability of ventilation capacity of hoods' and 'cleanliness of drainage'. On the basis of the findings of this study, we developed sanitary training posters, especially for small-scale restaurant operators. This could be an effective tool to educate food service employees on sanitary knowledge and principles and could be used to improve the existing sanitary conditions in Korean food service facilities.

The Effect of Freeze and Thaw for the Stabilized Soil Bottom Liners in the Landfill (폐기물 매립지 바닥층의 고화토 포설시 동결/융해 현상에 관한 연구)

  • Lee, Song;Lee, Jai-Young;Kim, Heung-Suck
    • 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.

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Evaluation of the stability of IgM and specific antibody response of sevenband grouper Epinephelus septemfasciatus for application of antibody-detection ELISA (항체검출 ELISA 적용을 위한 능성어 IgM의 안정성 및 특이 항체 반응 평가)

  • Kim, Chun-Seob;Jang, Min-Seok;Kim, Wi-Sik;Kim, Jong-Oh;Kim, Du-Woon;Kim, Do-Hyung;Han, Hyun-Ja;Jeong, Sung-Ju;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.335-342
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    • 2009
  • The stability of immunoglobulin M (IgM) on different serum storage conditions and specific antibody response were tested using the serum collected from sevenband grouper Epinephelus septemfasciatus by enzyme-linked immunosorbent assay (ELISA). To test the effect of storage temperature and duration, sevenband grouper antiserum against bovine serum albumin (BSA) was stored at -80, -20 or 4${^{\circ}C}$ for 1, 34, 61 or 119 days. In addition, to test the effect of repeated freeze-thawing condition, the anti-BSA fish serum was frozen at -20 and -80${^{\circ}C}$ and then thawn and frozen for 1, 5 or 10 times repeatedly. Consequently, no significant difference was found in ELISA optical density (O.D.) values of sera for the above mentioned storage conditions: different temperatures (-80, -20 and 4${^{\circ}C}$), durations of storage (1, 34, 61 and 119 days), and repeated thaw-freeze cycles (1, 5, and 10 times), indicating that IgMs of test fish were stable. The specific antibody response of sevenband grouper was observed after BSA-immunization of the test fish reared at 20 ${^{\circ}C}$ or 25${^{\circ}C}$. At the rearing temperature of 20${^{\circ}C}$, the specific antibody against BSA first appeared at 14 days and maximum antibody titer was observed between 21 and 28 days, while at the rearing temperature of 25 ${^{\circ}C}$, specific antibody appeared at 7 days and maximum antibody titer was observed between 14 and 21 days. In conclusion, the rearing temperature at 25${^{\circ}C}$ gave a faster and higher specific antibody response than at 20${^{\circ}C}$ and the specific antibody response maintained for approximately 2 months at 20℃ and 25${^{\circ}C}$.

The Evaluation of Various Conditions in the Cryopreservation of Primordial Germ Cells on Korean Native Chicken (Ogye) (한국재래닭(오계)의 원시 생식 세포의 냉동 보존에 있어서 여러 조건의 평가)

  • Kim, Hyun;Cho, Young Moo;Han, Jae Yong;Choi, Sung Bok;Byun, Mi Jeong;Kim, Young Sin;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Kim, Sung Woo
    • 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.