• Title/Summary/Keyword: frozen-thawed

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Comparative Study on the Viability of Frozen-thawed Primordial Germ Cells using Vitrification in Chicken Breed (초자화 동결법을 이용한 닭 품종간의 원시생식세포 동결성적의 비교)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Choi, Sung Bok;Ko, Yeoung Gyu;Do, Yoon Jung;Seong, Hwan Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.207-216
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    • 2013
  • This study was conducted to establish the method for preserving PGCs that enables long-term storage in liquid nitrogen for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of freeze-thaw treatment on viability of PGCs in chickens. PGCs were collected separately from a germinal gonad of an early embryo of 5.5~6 day (stage 28) of Isa brown, Korean Oge (KO), White Leghorn and Commercial breeds. PGCs separated from a germinal gonad of an early embryo of 5.5~6 day (stage 28) are suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and glycerol). The PGCs were then purified using magnetic activated cell sorting (MACS) method. The viability of PGCs after thawing was $87.4{\pm}0.4%$ and $89.4{\pm}0.2%$ with the 10% EG treatments with no significant difference between the Isa brown and Commercial breeds. The viability of PGCs after freeze- thawing was significantly higher for Isa brown ($87.4{\pm}0.4%$) and Commercial breeds ($89.4{\pm}0.2%$) than Korean Oge (KO) ($77.6{\pm}1.1%$) and White Leghorn ($76.2{\pm}0.9%$)(p<0.05) using 10% EG cryoprotectant. This study established a method for pre- serving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at agermplasm 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.

Consideration on Birth Sex Ratio in Human IVF-ET Program in Korea (인간의 체외수정 및 배아이식술에서 출생 성비에 대한 고찰)

  • Cha, Byung-Hun;Jun, Jin-Hyun;Lee, Jung-Ryeol;Choi, Young-Min
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.33-39
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    • 2010
  • Objective: Birth sex ratio (BSR) with human IVF-ET program is an interesting subject of social and scientific issue but very little information is available in Korea. This study was performed to assess the BSR with IVF-ET and to suggest the effective factors on the BSR. Methods: The national data from 1991~2008 were obtained from governmental Statistics Korea and the delivery data of human IVF-ET program on 2007 and 2008 were provided from the Ministry for Health, Welfare and Family Affairs. The BSR were statistically analyzed according to methods of IVF and to transferred embryos from fresh or frozen-thawed cycles. Results: The BSRs of Korean populations were over 1.10 up to 2002, and then it declined and maintained to 1.06 as balance BSR on 2007 and 2008. In human IVF-ET program, the BSRs were 1.07 on 2007 and 1.06 on 2008, respectively. Conventional IVF on 2008 showed the highest BSR as 1.10, and ICSI the lowest on 2008 as 1.01. There was no significant difference of BSRs related to the methods of in vitro fertilization and the feature of transferred embryos. Conclusion: The BSR of Korea showed 1.06~1.07 as normal and balanced state on 2007 and 2008 both general populations and human IVF-ET program. To the best of our knowledge, this is the first study to evaluate the BSR of human IVF-ET in Korea. There is a need to expand the further studies for national statistics and influencing factors on the BSR with IVF-ET.

Studies on Cryo-preservation of Registered Strains of Lentinula edodes (표고 등록균주의 초저온 보존에 관한 연구)

  • Ryu, Sung-Ryul;Bak, Won-Chull;Koo, Chang-Duck;Ka, Kang-Hyeon
    • Journal of Korean Society of Forest Science
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    • v.98 no.1
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    • pp.115-124
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    • 2009
  • New strain needs to maintain desirable characteristics for long term when it was bred, but in lapse of time it degenerates into a bad condition. Therefore the influence of temperature on the viability and survival rates of Lentinula edodes strains were examined after cryopreservation. Also, liquid nitrogen preservation for L. edodes has been proved to be one of the most reliable method. However, a mechanical damage of strain is inevitable during cryopreservation of the fungus because the fungus is very sensitive to stress of cooling rate in the freezing process. So we tried to find out state change of L. edodes with a programmable freezer. L. edodes strains were preserved at $-20^{\circ}C$, $-80^{\circ}C$ and $-196^{\circ}C$ for 50 days. At $-20^{\circ}C$, its mycelial growth became extinct. When thawed, the growth of mycelia which were preserved at $-80^{\circ}C$ was fastest. Attempts were made to investigate viability of L. edodes strains after freezing at $-80^{\circ}C$ and $-196^{\circ}C$, respectively. As the result, more than 90% showed high survival rate of strains tested at $-80^{\circ}C$ and $-196^{\circ}C$. Mycelial growth between apical and basal parts of colony after freezing preservation for 50 days was compared. At apical and basal parts, the survival rates showed 100% at $-80^{\circ}C$, but 98% and 94% at $-196^{\circ}C$, respectively. We confirmed that the ice crystal formation temperatures of L. edodes strains were $-6.0^{\circ}C$ for Sanlim 1, $-5.5^{\circ}C$ for the Sanlim 2, $-4.0^{\circ}C$ for the Sanlim 3 and $-15.5^{\circ}C$ for the Sanzo 302. These results indicated that L. edodes strains showed completely different responses to the ice crystal formation. We knew the fact that even the same species, especially L. edodes, they displayed completely different responses to the same freezing condition. Also, this has nothing to do with the connection between temperature type and freezing point. And a protocol was tried to minimize state change of L. edodes strains using programmable freezer when they are frozen, but it was not effective on them.

Effect of Cryopreservation by Slow and Rapid Freezing on the Sperm Motility Index, Viability and Morphology of Post-thaw Human Spermatozoa (인간 정자의 완만.급속 동결보존 방법이 융해 후 정자 운동성 지수와 생존율 및 정자 형태에 미치는 영향)

  • 김은국;김정욱;김형우
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.43-50
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    • 2003
  • The objective of this study was to investigate the effect of cryopreservation by slow and rapid freezing on the sperm motility index, viability and morphology of post-thaw human spermatozoa. After rapid freezing and thawing, sperm motility index was significantly higher (MOT:47.40$\pm$20.06%, VCL : 38.12$\pm$15.58 $\mu$m/s, VSL : 28.19$\pm$14.10 $\mu$m/s, VAP:33.64$\pm$15.15 $\mu$m/s, and HYP 2.77$\pm$2.71%) than slow freezing and thawing(MOT : 43.39$\pm$ 18.79%, VCL .33.91 $\pm$ 13.50 Um/s, VSL . 19.98$\pm$0.88 $\mu$m/s, VAP : 24.60$\pm$11.72 $\mu$m/s, and HYP . 1.33$\pm$1.57% ; P<0.05). But sperm Linearity(LIN) was significantly lower(28.83 $\pm$ 10.35) comparing to the slow freezing method(34.64 $\pm$ 11.36 ; P<0.05). On the other hand, significant difference were not observed MAD, WOB, DNC and DNM by slow and rapid frozen-thawed methods. After rapid freezing and thawing, sperm viability was lower(60 $\pm$ 2.2%) than slow freezing method(62 $\pm$2.1%) and sperm morphology was higher(46$\pm$7.7%) than that(44: 8.3). But there was no significantly These results indicate that rapid freezing method was positive effect of sperm cryopreservation in human.

Analysis of Massive Transfusion Blood Product Use in a Tertiary Care Hospital (일개 3차 의료기관의 대량수혈 혈액 사용 분석)

  • Lim, Young Ae;Jung, Kyoungwon;Lee, John Cook-Jong
    • The Korean Journal of Blood Transfusion
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    • v.29 no.3
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    • pp.253-261
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    • 2018
  • Background: A massive blood transfusion (MT) requires significant efforts by the Blood Bank. This study examined blood product use in MT and emergency O Rh Positive red cells (O RBCs) available directly for emergency patients from the Trauma Center in Ajou University Hospital. Methods: MT was defined as a transfusion of 10 or more RBCs within 24 hours. The extracted data for the total RBCs, fresh frozen plasma (FFP), platelets (PLTs, single donor platelets (SDP) and random platelet concentrates (PC)) issued from Blood Bank between March 2016 and November 2017 from Hospital Information System were reviewed. SDP was considered equivalent to 6 units of PC. Results: A total of 345 MTs, and 6233/53268 (11.7%) RBCs, 4717/19376 (24.3%) FFP, and 4473/94166 (4.8%) PLTs were used in MT (P<0.001). For the RBC products in MT and non-MT transfusions, 28.0% and 34.1% were group A; 27.1% and 26.0% were group B; 37.3% and 29.7% were group O, and 7.5% and 10.2% were group AB (P<0.001). The ratios of RBC:FFP:PLT use were 1:0.76:0.72 in MT and 1:0.31:1.91 in non-MT (P<0.001). A total of 461 O RBCs were used in 36.2% (125/345) of MT cases and the number of O RBCs transfused per patient ranged from 1 to 18. Conclusion: RBCs with the O blood group are most used for MT. Ongoing education of clinicians to minimize the overuse of emergency O RBCs in MT is required. A procedure to have thawed plasma readily available in MT appears to be of importance because FFP was used frequently in MT.

Investigation of Food Safety Attitude, Knowledge, and Behavior in College Students in Gyeonggi Region (경기도 지역 대학생의 식품 안전성에 대한 태도와 지식 및 행동 분석)

  • Kim, Ji-Myung;Hong, Seung-Hee
    • Journal of Food Hygiene and Safety
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    • v.33 no.6
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    • pp.438-446
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    • 2018
  • The purpose of this study was to investigate food safety awareness, knowledge, and behavior in college students, to provide basic data for the increase in food safety awareness. Data were collected from 252 college students in Gyeonggi region, using a self-administered questionnaire. In results of concern about food safety, subjects responded 3.48 of 5.00 and have knowledge about food safety education revealing significantly higher awareness and concern than subjects without knowledge about food safety education. Food safety awareness of distributed food was 2.55, considered unsafe. Among reasons in perceiving food as unsafe, 62.3% of subjects expressed distrust about safety relative food production. As for risk factors relative to food safety, subjects responded that the highest risk factor was food additives (2.35), followed by heavy metal (2.38) and endocrine disrupters (2.38). Correlation analysis resulting in risk factors for food had positive correlation with each other, heavy metal revealed highest correlation with pesticide residue (r = 0.674), than with endocrine disrupters (r = 0.672). Also, genetically modified food revealed high correlation with radiation irradiated food. Regression analysis demonstrated that concern about food safety significantly influenced pro-actively engaging in food safety education. Meanwhile, 63.5% of subjects correctly responded to food safety knowledge items. The item 'the heavy metals are contaminated the most, in the roots of vegetables' revealed the lowest correct answer rate (38.1%). In food safety behavior, the item 'always wash hands before handling food and meal's revealed 3.85, and subjects with awareness and concern about food safety education, responded in significantly higher numbers than subject without awareness and concern about food safety. The most neglected concern was relative to frozen food thawed at room temperature. Together, students recognize that distributed foods are unsafe, and students with awareness and concern about food safety education showed higher knowledge compared to without awareness and concern experience about food safety eduction. So, systematic education using accurate and objective data is required to reduce anxiety and raise the level of awareness and concern about food safety.

Effect of Sperm Preincubation Medium with Ascorbic Acid and/or Ferrous Sulfate on Porcine In-Vitro Fertilization (돼지의 체외수정시 Ascorbic Acid와 Ferrous Sulfate의 첨가하에서 정자 전배양의 영향)

  • Park, C.K.;Nam, H.S.;Lee, J.H.;Kim, I.C.;Cheong, H.T.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.24 no.3
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    • pp.255-262
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    • 2000
  • The aim of this work was to study the effects of ascorbic acid (Asc) and/or ferrous sulfate (Fe$^{2+}$) and spernatozoa preincubation on the in vitro fertilization in porcine. Porcine follicular oocytes matured in culture were inseminated with frozen-thawed boar spermatozoa preincubated for 0, 1, 2, 3, 4 and 5h. The penetration rates (37~51%) were not significantly different between durations of spermatozoa preincubation in medium with 0.1 mM Asc. The addition of 1.0 mM Fe$^{2+}$ during spermatozoa preincubation were not significantly affecting the penetration rates (41~56%). When spermatozoa were preincubated with Asc and Fe$^{2+}$, the penetration rates had a tendency to increase with time of spermatozoa preincubation, and were significantly (P<0.05) higher in spermatozoa preincubated with that than without Asc and Fe$^{2+}$ for 5 h. On the other hand, when spermatozoa were preincubated in fertilization medium without Asc and/or Fe$^{2+}$, the penetration rates were significantly (P<0.05) higher in medium with Fe$^{2+}$ than with Asc or Asc and Fe$^{2+}$ for in vitro fertilization. The rate of polyspermy in penetrated oocytes in medium with Asc and Fe$^{2+}$ decreased with the period of spermatozoa preincubation. Despite different culture conditions for spermatozoa preincubation, no differences were observed in polyspermy rates in the presence of Asc and/or Fe$^{2+}$ These results indicate the advantage of preincubating spermatozoa with Asc and Fe$^{2+}$ and an addition of Fe$^{2+}$ during in vitro fertilization with spermatozoa preincubated maintain penetration potential without increased polyspermy rates on in vitro fertilization in porcine oocytes.on in porcine oocytes.

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