• Title/Summary/Keyword: 'Frozen 2'

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Studies on Embryo Transfer in Rabbit ―II. The viability of deep-frozen embryos at different developing stages― (가토의 수정란이식에 관한 연구 ―II. 동결융해난자의 발육단계별 생존성-)

  • 김정익;양부근;남상헌;고광두
    • Korean Journal of Animal Reproduction
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    • v.7 no.1
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    • pp.19-23
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    • 1983
  • Present studies were conducted to investigate the developmental stage and the location of embryos in the reproductive tract at various times after ovulation, the morphologically normal after thawing of embryos preserved in liquid nitrogen, and the survival after transferring frozen-thawed embryos. The results obtained were as follows: 1. Embryo stage and location in the reproductive tract after hCG administration. For the investigation of embryo stage and location in the reproductive tract after ovulation, rabbits were laparotomized at 24, 40, 48, 72 and 120 hrs post hCG injection, simultaneously with mating. the oviducts and uteri were flushed out with PBS medium containing 50% rabbit serum, respectively. 1) Most of embryos was remained in the oviduct within 48 hrs, with the lapse of time, embryos were started to move to uterus and shifted in uterus at 72 hrs after hCG injection. 2) The representatives of embryos stage collected at 24, 40, 48, 72 and 120 hrs were 1-cell(60.4%), 8-cell to early morula (52.3, 39.3%), late blastocyst (95.5%) stages, respectively. 2. Morphological normality and survival of the frozen-thawed embryos. For the evalution of the quality and viability on the frozen-thawed embryos, immediately after thawing, embryos were assessed by morphologically normal under a dissecting microscope, and a further test of frozen-thawed embryos was made by transferring the morphologically normal embryos to the uteri of recipient rabbit induced pseudopregnancy by the injection of hCG at the time of hCG injection in donor rabbits. 1) The propotions of embryos which a, pp.ared morphologically normal was higher when 8-cell (85.7%) and morula(90.5%) were used for freezing than when 4-cell (66.7%) and blastocyst (75.8%) were used. 2) Preganacies were observed at Day 15 after transfer of frozen-thawed 8-cell (7/13), morula (19/42) and blastocyst (3/19) but not after transfer of embryos at 4-cell stage.

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Effect of Ethylene Glycol and Antioxidant Combination on Function of Frozen-thawed Spermatozoa in Korean Jeju Black Bull (Ethylene glycol과 항산화제가 제주흑우 동결정액에 미치는 영향)

  • Ko, Min-Hee;Seo, Jong-Pil;Kang, Tae-Young
    • Journal of Veterinary Clinics
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    • v.34 no.2
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    • pp.156-160
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    • 2017
  • We investigated the effect of ethylene glycol and antioxidants such as taurine, hypotaurine and trehalose with extenders during cryopreservation of Korean Jeju Black Bull spermatozoa. The cryopreservation of freshly collected spermatozoa was conducted with four different conditions. As a control, spermatozoa were cryopreserved with Tris egg-yolk extenders added 5% ethylene glycol (EG). Taurine (20 mM), hypotaurine (20 mM) and trehalose (20 mM) were individually added into tris egg-yolk extenders with 5% EG. After thawing of frozen spermatozoa with four different conditions, sperm viability, motility, acrosomal integrity, and membrane integrity were investigated. The significant (p < 0.05) improvement of sperm viability showed in all antioxidant treated thawed spermatozoa (taurine; $68.1%{\pm}4.4$, hypotaurine; $69.2%{\pm}6.7$ and trehalose; $68.0%{\pm}4.4$) when compared to control ($63.4%{\pm}5.6$). Neither positive nor detrimental effects of three antioxidants were shown sperm motility after thawing. The results of hypo-osmotic swelling test showed that the membrane integrity of taurine, hypotaurine or trehalose treated thawed spermatozoa ($64.1%{\pm}5.4$, $61.5%{\pm}3.7$ and $59.0%{\pm}4.0$, respectively) had significantly (p < 0.05) higher rate of the swollen sperm compared to control ($53.7%{\pm}9.7$). Hypotaurine treated frozen-thawed spermatozoa had siginificantly higher (p < 0.05) F pattern ratio than taurine, trehalose and control treated frozen-thawed spermatozoa. Trehalose added frozen-thawed spermatozoa had significantly higher (p < 0.05) acrosome reaction pattern ratio than taurine and hypotaurine added frozen-thawd spermatozoa. In this study, we found that antioxidants such taurine, hypotaurine and trehalose treatments during cryopreservation process could reduce damage of spermatozoa of Korean Jeju Black Bull and improved sperm capability of fertilization.

Changes in Range of Motion after Intra-Articular Corticosteroid Injection in Frozen Shoulder: A Retrospective 3-Month Follow-Up Study (동결견 환자에서 관절강내 스테로이드 주사 후 관절가동범위의 변화)

  • Cho, Hyoung Jun;Yoon, Seung-Hyun;Kim, Minchul;Kim, Dae Hwan
    • Clinical Pain
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    • v.18 no.2
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    • pp.76-81
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    • 2019
  • Objective: To examine (1) the degree of reduction of passive range of motion (PROM) on the affected side compared to that on the unaffected side and (2) the degree of increase in PROM following intra-articular corticosteroid injection (IACI) in patients with frozen shoulder. Method: The medical records of 120 patients with frozen shoulder were retrospectively reviewed. PROM of the unaffected and affected shoulder (flexion, extension, abduction, internal rotation, external rotation) was compared, and changes in PROM of the affected shoulder after a single IACI (triamcinolone 20 mg) were evaluated after 12 weeks. Results: At the time of diagnosis, PROM of the affected shoulder was most limited in external rotation, followed by internal rotation, abduction, extension, and flexion, compared to that of the unaffected shoulder. Compared to before IACI, PROM of external rotation demonstrated the greatest increase compared to all the other movements after IACI. Conclusion: Limitation in PROM of the frozen shoulder at the time of diagnosis was greatest for external rotation. Moreover, external rotation experienced the greatest improvement after IACI. Our findings should help to further clarify the clinical characteristics of frozen shoulder, aid in its diagnosis, and allow the prediction of the effects of IACI.

Changes in the Properties of Myofibrillar Proteins from Broiler Meat during Cold and Frozen Storage (육계육의 냉장 및 동결저장 중 근원섬유 단백질의 특성 변화)

  • 박창식;문윤희
    • Korean Journal of Poultry Science
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    • v.14 no.2
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    • pp.137-143
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    • 1987
  • This study was carried out to compare the changes in the extractability, biological activity, and solubility of myofibrillar proteins and actomyosins during storage period at 4$^{\circ}C$ and -20$^{\circ}C$in pectoral. and leg muscle of broiler meat. 1. The results obtained are as fellows ; The extractabilities of myofibrillar proteins in pectoral and leg muscle were increased gradually to 7-days during storage at 4$^{\circ}C$ and decreased slightly during frozen storage at -20$^{\circ}C$. The extractabilities of actomyosins in pectoral and legmuscle were not greatly changed during cold storage and decreased gradually during frozen storage. 2. The Ca$\^$2+/-ATP ase activities of myofibrillar proteins in the both muscles were not greatly changed to 7-days during cold storage, and in the case of frozen storage, those were highest on the 2nd week, thereafter decreased with storage period. The Ca$\^$2+/-ATPase activities of actomyosins in pectoral and leg muscle were decreased sightly only frist day during cold storage and decreased gently during frozen storage. 3. Myofibrillar proteins in the both muscles were solubilized completely at 0.20M KCl in fresh meat, at 0.25M (pectoral) and 0.30M KCl (leg) in the cold storage, and at 0.30M KCl in the frozen storage. Actomyosins of both muscles were solubilized completely at 0.40M KCl in fresh meat, cold and frozen storage.

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Survival and In Vitro Development Rate of Frozen Mouse Embryos in Various Cryoprotectants (항동해제에 따른 생쥐 동결수정란의 생존율및 체외발달율)

  • Cha, Sang-Hun;SunWoo, Jae-Gun;Park, Hyo-Suk;Lee, Im-Soon;Cho, Tai-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.2
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    • pp.167-172
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    • 1990
  • This study was carried out to clarify the effects of various kinds of cryoprotectants which were frequently used in freezing embryos of domestic animals on the survival of frozen-thawed mouse embryos. Mouse embryos were collected by hyperstimulation induction of ICR mouse. The samples were slowly cooled ($l^{\circ}C/min$) to temperatures between $-7^{\circ}C$ and $-30^{circ}C$ before direct transfer to liquid nitrogen ($-196^{\circ}C$) and thawed rapidly ($-500^{\circ}C$/min). As cryoprotectants, Glycerol, DMSO, Ethylene glycol and Propylene glycol were used and applied each 2 cell, 8 cell, morula in embryo stage. After normal mouse embryos developed to blastocyst by in vitro culture, we observed recovery rate and developing rate of embryos at thawing. The results obtained in these experiments were as follows : 1. The in vitro development rate from the frozen-thawed 2 cell embryos to the blastocyst were 67.7% in ethylene glycol, 65.7% in Propylene glycol, 55.2% in glycerol and 50.0% in DMSO respectively. 2. The in vitro development rate from the frozen-thawed 8 cell embryos to the blastocyst were 83.6% in DMSO, 75.7% in glycerol, 52.2% in propylene glycol respectively. 3. The in vitro development rate from the frozen-thawed morula to the blastocyst were 84.2% in glycerol, 80.0% in DMSO, 66.6% in propylene glycol and 55.2% in ethylene glycol respectively.

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Efficacy of Frozen-Thawed ET in Patients with Old Age or Non-Pregnant in Fresh ET Cycles (고령 환자와 신선주기 배아이식에서 임신에 실패한 환자에서 동결-융해 배아이식의 효용성)

  • Choi, Su Jin;Lee, Sun Hee;Song, In Ok;Koong, Mi Kyoung;Kang, Inn Soo;Jun, Jin Hyun
    • 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.

Changes of Fatty Acid Composition in Shank During Heating Time and Frozen Storage (사태의 가열시간 및 냉동저장에 따른 지방산 조성 변화)

  • Kim, Kyung-Ae
    • Korean journal of food and cookery science
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    • v.2 no.2
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    • pp.8-15
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    • 1986
  • This study was carried out to investigate changes of the lipid contents and the fatty acid composition in shank during heating time and frozen storage. 1. The total lipid contents of raw shank were about 3.57% and decreased stepwise during heating time 30, 60, 90 min and frozen storage(24hrs) The contents of neutral lipid, glycolipid and phospholipid were 70.71%, 6.36%, and 22.93% in raw shank, and neutral lipid contents were decreased, whereas Phospholipid contents were increased according to heating tide. In frozen storage, neutral lipid and glycolipid contents were increased, while phospholipid contents were decreased. 2. Lipids of shank possessed about 8 kinds of fatty acid as the constituent by gas-liquid chromatography analysis. The main fatty acids were oleic acid, palmitic acistearic acid and linoleic acid: the fatty acids of total lipids in raw shank were 43.48% of oleic acid, 23.13% of palmitic acid,12.00% of stearic acid and 6.75% of linoleic acid. Also the fatty acids were 43.32% of oleic acid, 23.26% of palmitic acid, 9.30% of stearic acid 2.15% of linoleic acid in neutral lipid, 22.63% of oleic acid, 8.44% of palmitic acid, 11.98% of stearic acid, 27.01% of linoleic acidin glycolipid, 39.38% of oleic acid, 15.89% of palmitic acid, 15.55% of stearic acid, 17.49% of linoleic acid in phospholipid. 3. The fatty acid pattern of total lipid, neutral lipid, glycolipid and phospholipid was not any changes, whereas there was a difference in the fatty acid contents: palmitic acid and stearic acid of total lipid were decreased in the 30 min and 60 min heating but increased in 90min heating, and linoleic acid of neutral lipid was increased stepwise during heating time and frozen storage. Also palmiict acid of glycolipid was increased gradually and linoleic acid in heating time 30, 60 min was higher than 90 min and frozen storage. Among fatty acids in phoapholipid, oleic acid was increased during heating time, while decreased in frozen storage, and linoleic acid was not any change but linolanic acid was increased. UFA/SFA of phospholipid was the highest when heating time was 60 min. From above results, it was found that when heating time was 60 min beneficial nutritionally, comparing with changes of fatty acid composition according to the heating time aid frozen storage.

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The Comparison and Distribution of Temperatures Established in Display Stands and Food Surfaces for Cold and Frozen Foods in large Discount Stores in Korea (대형할인매장에서의 냉장.냉동식품 판매대의 설정온도와 판매식품 표면온도 비교 및 온도분포 분석)

  • Choi, Moon-Sil;Choi, Jeong-Ae;Kim, Mee-Hye;Bahk, Gyung-Jin
    • Journal of Food Hygiene and Safety
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    • v.26 no.4
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    • pp.308-314
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    • 2011
  • This study surveyed and compared the temperatures established in display stands and food surfaces for cold and frozen foods in large discount stores in Korea. The temperatures established in display stands for cold food ranged with $3.5{\pm}1.8^{\circ}C$ as mean, minimum and maximum were $0^{\circ}C$ and $7^{\circ}C$. However, the surface temperatures of cold food on sale ranged with $10.7{\pm}2.9^{\circ}C$ as a mean, minimum $4.6^{\circ}C$ and maximum $18.4^{\circ}C$. Totally, the surface temperature of cold food on sale was $7.2^{\circ}C$, as a mean, higher than established in display stands for cold food in large discount stores in Korea. 53% of the surveyed cold foods were more than $10^{\circ}C$ in surface temperature and only 47% was less than $10^{\circ}C$. The differences between temperatures were lowest in fruits, salads and vegetables, but highest in milk products. On the other hand, the temperatures established in display stands for frozen food showed a range with $-20.7{\pm}1^{\circ}C$ as a mean. However, the surface temperatures of frozen food on sale showed a range with $-15.4{\pm}5^{\circ}C$ as a mean, minimum $-28^{\circ}C$ and maximum $-4.6^{\circ}C$ (included defrosting). The surface temperatures of frozen food, frozen meats, frozen processed foods and ice creams were $-13.8^{\circ}C$, $-15.9^{\circ}C$, and $-16.8^{\circ}C$, respectively. Only 32.3% of surveyed frozen foods showed less than $-18^{\circ}C$ in surface temperature. In conclusion, the temperatures established on cold and frozen food display stands were less than those of cold and frozen food surfaces on sale. There was also much variation in food surface temperatures during cold and frozen food storage and sales. Therefore, a temperature management system technology use at the distribution level for cold and frozen foods will be developed.

Improved pregnancy rate and sex ratio in fresh/frozen in vivo derived embryo transfer of Hanwoo (Bos taurus coreanae) cows

  • Jihyun Park;Wonyou Lee;Islam M. Saadeldin;Seonggyu Bang;Sanghoon Lee;Junkoo Yi;Jongki Cho
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.779-791
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    • 2023
  • This study aimed to assess the effects of embryonic developmental stage, quality grade, and fresh or frozen/thawed conditions on the pregnancy rate and sex ratio of live offspring in Hanwoo (Bos taurus coreanae) cows. The quality and developmental stage of in vivo-derived (IVD) transferred embryos were evaluated using the standard criteria of the International Embryo Technology Society. The recipient cows were synchronized using conventional (estradiol benzoate and progesterone) protocols before embryo transfer. Embryos were transferred to 297 cows, and pregnancy was monitored for 60-70 days after embryo transfer. The pregnancy rates of fresh and frozen/thawed embryos were 56.90% and 52.49%, respectively. Pregnancy rates varied according to embryo quality (56.18% for grade 1 vs. 36.67% for grade 2). Pregnancy rates also varied by developmental stage and cryopreservation (67.86% vs. 63.49% for stage 4-1, 64.00% vs. 54.72% for 5-1, and 50.00% vs. 47.83% for 6-1, in fresh embryos vs. frozen/thawed embryos, respectively). For stage 7-1, the pregnancy rates were 72.73% for fresh embryos and 20.00% for frozen/thawed embryos. In 66 fresh embryos, the sex ratio of live offspring was 5:5, whereas it was 4(female):6(male) for frozen/thawed embryos among the 95 frozen/thawed embryos. The miscarriage rate was approximately 3% higher for frozen/thawed embryos than for fresh embryos (18.1% for fresh vs. 21.1% for frozen). Seasonal fertility rates were 33.3% in spring, 55.67% in summer, 52.8% in autumn, 60.0% in winter. The following male-to-female ratios were observed in different seasons: 6.7:3.3 in spring, 4.0:6.0 in summer, 5.5:4.5 in autumn, and 3.3:6.7 in winter. The current data revealed no significant differences in pregnancy rates between fresh and frozen/thawed IVD embryos. However, there was a lower pregnancy rate with advanced-stage frozen/thawed embryos (stage 7-1). The current study provides comprehensive results for the better optimization of embryo transfer in Hanwoo cattle to obtain the desired fertility rate, pregnancy rate, and sex ratio of calves. These results provide important insights into the factors that influence the viability and success of IVD embryo transfer in Hanwoo cows and may have practical applications for improving breeding programs and reducing production costs.

Effects of Thawing Conditions on the Viability and Acrosomal Morphology of Cryopreserved Boar Semen (동결보존한 돼지정액의 융해조건이 정자의 생존율과 첨체변화에 미치는 효과)

  • 정영호;서경덕;김광식;심금섭;이장희
    • Journal of Embryo Transfer
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    • v.14 no.2
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    • pp.131-137
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    • 1999
  • This experiment was carried out to investigate the effects of osmolarity of thawing diluents, seminal plasma added in thawing diluents on the sperm viability and the effects of thawing temperature, the temparature of the thawing diluents on the sperm viability and acrosomal morphology of boar spermatozoa by the straw method. The result obtained were summarized as follows: 1. The sperm viablilty after thawing of the frozen semen was shown greater in the high osmolarity(392~492mOsm) than low osmolarity(300mOsm) in thawing diluent. The added levels of seminal plasma in thawing diluent did not affect the viability of frozen-thawed boar semen. 2. In terms of thawing temperature, the sperm viability was shown higher in the frozen semen thawed at 5$0^{\circ}C$ for one min. (p<0.01) than those thawed at 2$0^{\circ}C$ or 37$^{\circ}C$ for one min. The sperm viability was not significant at the diluent temparature of 2$0^{\circ}C$or 37$^{\circ}C$ after thawing: but the sperm viability was higher in thawing diluent at 2$0^{\circ}C$ than in that at 37$^{\circ}C$. However, the effects of thawing temperature and diluent solution on normal acrosomal rate were not significant. 3. Cleavage rates of oocytes fertilized with frozen semen were 46.4% and 43.3%, respectively, which were thawed at 5$0^{\circ}C$ for one min. and then diluted in mBTS medium at 2$0^{\circ}C$or 37$^{\circ}C$. To sum up, the sperm viability was shown greater at the high of thawing diluents of frozen boar semen. In terms of thawing conditions, the sperm viability was shown greater, when semen was thawed at a high temperature for a short time and then diluted at the same temperature as that in the straw.

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