• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.22 no.3
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• pp.262-269
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• 2019

Purpose: As the importance of breastfeeding has been reinforced, human milk is often stored for practical reasons. Therefore, we evaluated optimal storage and processing methods for human milk from a nutritional standpoint. Methods: Human milk samples were collected between June 2017 and February 2018. Also, data about maternal information were collected. Human milk was analyzed for macronutrients and caloric content. The samples were subdivided into groups for nutrient analysis. The control group (fresh milk) was not stored or processed. The other groups (9 groups) consisted of samples analyzed based on different storage temperatures (room temperature, refrigerated, frozen), defrosting methods (bottle warmer, room temperature thawing, microwave oven), and storage period (1 week, 1 month, 2 months) and compared with the control group. Results: There was no statistically significant difference in the nutrient content of human milk among the collected samples. A significant change in the content of macronutrients in milk samples was observed under storage condition at different temperatures for 1 week with subsequent thawing with bottle warmer compared to fresh milk. Under storage at $-20^{\circ}C$ for 1 week with subsequent thawing with different defrosting methods, a significant change in the content of macronutrients in milk samples was observed compared to fresh milk. After storage at $-20^{\circ}C$ for different periods and thawing with a bottle warmer, a significant change in macronutrient content in milk samples was observed compared to fresh milk regardless of the storage period. Conclusion: Unlike previous guidelines, changes in macronutrient content in milk samples were observed regardless of the method of storing and thawing. Apparently, it is proposed that mothers should feed fresh human milk to their babies without storing.

• Clinical and Experimental Reproductive Medicine
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• v.39 no.4
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• pp.153-160
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• 2012

Objective: The aim of this study was to compare vitrification optimization of mouse embryos using electron microscopy (EM) grid, cryotop, and thin plastic strip (TPS) containers by evaluating developmental competence and apoptosis rates. Methods: Mouse embryos were obtained from superovulated mice. Mouse cleavage-stage, expanded, hatching-stage, and hatched-stage embryos were cryopreserved in EM grid, cryotop, and TPS containers by vitrification in 15% ethylene glycol, 15% dimethylsulfoxide, 10 ${\mu}g/mL$ Ficoll, and 0.65 M sucrose, and 20% serum substitute supplement (SSS) with basal medium, respectively. For the three groups in which the embryos were thawed in the EM grid, cryotop, and TPS containers, the thawing solution consisted of 0.25 M sucrose, 0.125 M sucrose, and 20% SSS with basal medium, respectively. Rates of survival, re-expansion, reaching the hatched stage, and apoptosis after thawing were compared among the three groups. Results: Developmental competence after thawing of vitrified expanded and hatching-stage blastocysts using cryotop and TPS methods were significantly higher than survival using the EM grid (p<0.05). Also, apoptosis positive nuclei rates after thawing of vitrified expanded blastocysts using cryotop and TPS were significantly lower than when using the EM grid (p<0.05). Conclusion: The TPS vitrification method has the advantages of achieving a high developmental ability and effective preservation.

• Korean Journal of Poultry Science
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• v.46 no.4
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• pp.223-234
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• 2019

This study investigated the effects of thawing methods and storage time on the quality of frozen duck meat. Meat was obtained from eight-week-old Korean native ducks (average weight=2.8 kg). Seventy-two samples were divided into eight treatments (three replicates/treatment, three samples/replicate) with 2 × 4 factorial arrangement based on two thawing methods (under running water at 12℃ for 3 h and in a refrigerator at 5℃ for 24 h) and four storage times (1, 3, 6, and 12 months). CIE b^* was significantly different among different storage time treatments, reaching its lowest after 6 months (P<0.05). Cooking loss did not differ between storage times; however, it was significantly lower following application of the fast thawing treatment (P<0.05). Water-holding capacity of meat stored for one month was highest compared to that of meat stored for a longer period (P<0.05). Additionally, there were significant differences based on storage time in γ-linoleic acid (C18:3n6) and eicosenoic acid (C20:1n9) contents (P<0.01), as well as in protein contents (P<0.05). Palmitoleic acid (C16:1n7) typically decreased after three months of storage; however, this decline was not significant compared to other storage times. Essential amino acids contents, except methionine, were significantly difference at six and 12 months of storage (P<0.05). Similarly, non-essential amino acid contents, except tyrosine, were significantly different among storage periods (P<0.05, P<0.01). Alternatively, there were no significant differences in the chemical composition, fatty acid content, or amino acid content based on the thawing method.

• Proceedings of the Korea Concrete Institute Conference
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• 2008.04a
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• pp.589-592
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• 2008

This study was conducted to assess the durability of Chloride-inhibiting Low-Heat Cement while being subjected to freezing-thawing during winter seasons. Although durability varies slightly depending on the conditions of the jobsite, frost damage to concrete resulting from repeated freezing and thawing over the course of seasonal changes is the leading cause behind lowered concrete durability. in addition, concrete that has been subjected to freezing and thawing during the winter season develops a significant amount of expansive force at the core and begins to exhibit signs of damage, such as cracking, peeling, and detachment from the aggregate. Therefore, this study fabricated test specimens using a Chloride-inhibiting Low-Heat Cement(CLC) and the widely used blast furnace slag cement(BFS) and Ordinary Portland Cement(OPC) with water-to-cement ratios of 35%, 40% and 45%, respectively, to assess the durability index of the CLC as per resistance to freezing-thawing. The specimens were then tested using the KS F 2456 method (Testing method for resistance of concrete to rapid freezing and thawing) to measure the dynamic modulus of elasticity. The dynamic modulus of elasticity measurements were then used to derive the durability indices. By comparing the durability indices, it was confirmed that CLC, BFS, and OPC all had superior durability.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.119-124
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• 2002

This study was carried out to investigate the effects of packing materials of frozen boar semen to improve reproductive performance efficiency in pig. Boars were raised at Swine Artificial Insemination Center in National Livestock Research Institute, Sunghwan, Chungnam, Korea. We compared packing protocols for frozen boar semen among 5$m\ell$ maxi-straw, 5$m\ell$ cryogenic-vial, and aluminum-pack. Cryogenic-vial packing material showed similar sperm characteristics compared with maxi-straw packing material when the sperm was frozen above 15cm from liquid nitrogen and thawed at 52$^{\circ}C$ for 190 seconds. We investigated different thawing times to find out the optimal condition of freezing and thawing protocol with cryogenic-vial. Freezing above 15cm from liquid nitrogen and thawing at 52$^{\circ}C$ for 190 seconds were the optimal protocol compared with 120 and 150 seconds. However, normal acrosome rates did not show any differences among thawing times. Post-thawing results of maxi-straw in water at 52$^{\circ}C$ for 45 seconds had better total motility and curve linear velocity than those of cryogenic-vial in water 52$^{\circ}C$ for 190 seconds. However, there were no differences on straightness and normal apical ridge of sperm between maxi-straw and cryogenic vial. Non-return rate, farrowing rate and litter size of sows inseminated with frozen boar semen of commercial farms were higher in the maxi-straw than cryogenic-vial, but there were no significant differences between maxi-straw and cryogenic-vial. In conclusion, there were no significant differences between maxi-straw and cryogenic-vial and so, we may replace cryogenic-vial packing method instead of maxi-straw packing method by improvement of freezing and thawing rate.

• Food Science and Preservation
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• v.18 no.3
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• pp.302-309
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• 2011

This study was performed to analyze the physicochemical and sensory properties of Acanthopanacis cortex and Aralia elata according to their blanching conditions and thawing methods. In terms of their Hunter colors, the A. cortex and A. elata that were blanched without adding salt to them for 7 min and 4 min, respectively, had the highest L values. The chlorophyll content (148.7 mg%) of A. cortex that was blanched with 1% salt for 4 min was higher than those of the other samples, and the chlorophyll content of A. elata was 32.4 mg% when it was blanched for 4 min without salt addition. The sensory test results showed the highest overall preference for the sample that was blanched without salt for 4 min among all the samples. The Hunter color of A. cortex did not significantly differ with different thawing methods, but the value of A. elata that was thawed in a microwave oven was higher than those of the other samples. The chlorophyll contents of A. cortex and A. elata that were thawed in a microwave oven were the highest among all the samples. As for the overall preference for the samples according to the thawing method, A. cortex and A. elata scored highest in the case of thawing at $25^{\circ}C$ and in a microwave oven, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.143-152
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• 1992

The present experiments were focussed to modify a short slow-cooling protocol used for freezing of early stage embryo(Testart et al., 1986) and also to apply the modified method for the cryopreservation of hamster oocytes with Zona or without. The protocol was modified by changing the 4-step equilibration into 1-step and the 1-step thawing into 2-step. The oocytes were added in 1.5M PROH and 0.1M Sucrose, seeded at $-7^{\circ}C$, slow cooled($0.3^{\circ}C$/min) to $-30^{\circ}C$ before plunging to $-196^{\circ}C$. The oocytes were thawed at $23-25^{\circ}C$ air(20sec/150sec) and/or 33-35 water(10sec). The survival of the frozen-thawed oocytes was determined by morphologic criteria and their fertilizing ability was also estimated by Sperm Penetration Assay(SPA) system(Chang et al, 1990) using fertile men semen sample. One-step equilibration showed slightly higher survival rate(83.9% vs. 71.0%) and fertilization rate(83.9% vs. 71.0%) compared with four-step(p>0.05). And two-step thawing(air & water exposing) of oocytes frozen after 1-step equilibration showed significantly higher survival rate(96.3%) than one-step thawing at air(85.2%) or water(65.0%) only(p<0.05). Therefore, by the modified method(l-step equilibration & 2-step thawing), Zona-intact(ZI) and Zona-free(ZF) oocytes were frozen and thawed. ZI-oocytes showed significantly higher survival rate(95.4%, 308/323 vs. 67.6%, 240/355) than ZF-oocytes(P<0.01). But the survival of ZF-oocytes was as high as ZI-oocytes in fourteen of twenty-four replicates. ZI-oocytes was also significantly higher fertilization rate($92.4{\pm}8.9%$ vs. $63.7{\pm}18.5%$) and higher mean number of penetrated sperm($6.2{\pm}4.2$ vs. $3.9{\\pm}3.3$) than ZF-oocytes, but not higher than control(fresh oocytes;$99.3{\pm}2.4%$, $8.4{\pm}4.2$)(P<0.001). Conclusively, this modified method will contribute to freeze effectively the hamster oocytes for simplifing of the logical consideration of performing SPA and also to freeze the human and other animal oocytes.

• Journal of Embryo Transfer
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• v.4 no.1
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• pp.1-13
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• 1989

This paper reviews the most important steps that have generated consistent progress in principles and developmental progress of embryo cryopreservation, and also study on freezing procedure and its application by conventional method and current improved method for freezing procedure and its appilcation of embryo cryopreservation in farm animals. Four were of particular interest: 1.The transport of water across the ccli membrane (zona pellucida) during freezing and thawing accordinglyplays a role in determing whether the celi survives. This movement of water is controlied mainly by extracellular phase changes and by the nature and concentration of any cryoprotective agent present. Therates of cooling, freezing and warming, and the intervals over which they are applied are further decisi've factors in determining whether a cryopreservation procedure allows survival after thawing. 2.The first successful deep freezing experiments with sheep morula and blastocysts during the seventies were based on the early procedures used for mouse embryos.Current research during the eighties is developed with the aim of simplifying and improving current procedures such as one-step dilution and rapid or ultra-rapid cooling by using the model of laboratory animals. 3.The conventional method for the embryo cryopreservation is described. An alternative to this method which may result in high survival and also in reducing of the freezing and thawing time is done by combing a permeable cryoprotectant such as glycerol, DMSO or propanediol and a non-permeable compound such as sucrose, trehalose, raffinose or lactose. 4.Finally a different approach to the preservation of embryos, named vitrification, is introduced. This procedure depends upon the ability of concentrated solutions of cryoprotective agents such as glycerol and propanediol to supercool to very low temperature (-196$^{\circ}C$) during rapid cooling before solidifying without formation of ice. However, more complete data are necessary for successful vitrification of blastocysts.

• Journal of Embryo Transfer
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• v.23 no.1
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• pp.19-24
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• 2008

The purpose of this study was to compare the efficiency of slow freezing with that of vitrification method for the cryopreservation of human embryos. Human embryos were derived from in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and the mixed solution of propanedial (1.5, 1.0, 0.5M PROH) and sucrose (0.1M), ethylene glycol (7.5, 15%), dimethyl sulfoxide (7.5, 15% DMSO), sucrose (0.5, 1.0M) and SPS (Serum Protein Substitute) was used for a cryoprotectant for slow freezing and vitrification solution, respectively. Rates of recovery after thawing, morphological normality, post-thaw viability, arrest, morphological abnormality and preimplantation development were compared between two protocols. After freezing-thawing, recovery and survial rate of slow freezing was (88.6% and 73.4%), whereas vitrification was (99.2% and 96.2%) (p<0.05). The arrest rate of slow freezing was significantly lower compared with those of vitrification(8.7% vs 29.9%) (p<0.05). Preimplantation development to the 2-cell (83.8% vs 67.7%), 4-cell (69.0% vs 47.2%) and 8-cell (62.4% vs 37.8%) stages 24, 48 and 72 h after thawing, respectively, were higher in the slow freezing than the vitrification. After slow freezing and vitrification of human embryo at 2-8cell stage, the rate of recovery rate, survival rate and partial damage rate were 92.0% vs 100%, 80.4% vs 96.2% and 52.2% vs 19.0%, respectively. And partial damage rate was significantly lower than those of slow freezing method (p<0.05). These results demonstrate that a slow freezing using PROH is more efficient than a vitrification for cryopreserving the human zygotes, although the vitrification yielded better recovery, survival and partial damage of frozen-thawed 2-8 cell stage embryos than slow freezing method.

• Journal of Embryo Transfer
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• v.7 no.2
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• pp.81-88
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• 1992

Cryopreservation of mouse embryos biopsied at 4-cell stage was investigated by ultrarapid freezing. Four-cell embryos were obtained from ICR mice on 55h after hCG injection. Zona pellucida of the embryos were partially dissected with a cutting pipet, and then single blastomeres were biopsied from the embryos followed by incubation in $Ca^2$+ and $Mg^2$+-free M16 medium for 30min. Biopsied embryos cultured for lh or 15h were frozen by ultrarapid freezing method using 3M DMSO or 5M glycerol as a cryoprotectant, respectively. The developmental rate of biopsied embryos after ultrarapid freezing and thawing to blastocysts was 81 % in the group of biopsied embryos cultured for lb and 98% in the group of biopsied embryos cultured for 15h, respectively. When biopsied embryos after ultrarapid freezing and thawing were transferred to the uteri of pseudopregnant recipients, normal live young were born. These results suggest that this freezing method can efficiently cryopreserve biopsied mouse embryos.