• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.75-75
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• 2001

유리화 동결법은 동결중 ice crystal의 형성이 이루어지지 않으므로 난자의 동결보존을 위한 유용한 동결방법이다. 이전의 연구에서 마우스의 난자를 ethylene glycol과 electron microscope grid를 이용한 유리화 동결법으로 동결 융해한 결과 기존의 slow freezing 방법에서보다 높은 생존율과 배발달율이 나타남을 관찰하였다. 그러나 동물과 인간 난자를 이용한 연구를 통하여 난자의 경우 동결 융해후 염색체에 부착되어 있는 미세소관인 방추사가 온도의 변화에 매우 민감하여 염색체 이상성이 증가되는 것이 보고되었다. 이에 본 연구에서는 성숙난자를 유리화동결법에 의해 동결 융해후 난자의 염색체와 방추사의 이상성이 증가되는지 알아보고자 본 실험을 수행하였다. ICR mouse의 성숙란을 채취하여 연구목적에 따라 fresh한 대조군과 동결음해 시킨 실험군으로 분류하였다. 동결방법은 난자를 1.5 M ethylene glycol (EG)에 2분 30초간 노출 시킨후 5.5 M EG와 1M sucrose가 첨가된 동결액에 20초간 노출시킨 후 Grid에 난자를 부착시킨 후 직접 액체질소에 침지하여 동결하였다. 동결후 난자는 5단계로 융해를 실시한 후 생존된 난자는 tubulin 항체를 이용한 immunostaining 방법으로 방추사의 이상성을 관찰하였고, 염색체는 난자를 고정 후 10% Giemza로 염색 후 염색체의 수적인 이상성을 관찰하였다. 염색체 분석결과 염색체 이상 빈도는 대조군의 경우 19.6%, 동결융해군은 32.8%로 관찰되었다. 또 방추사의 이상빈도는 대조군의 경우 20.2%, 동결 융해군은 32.3%로 관찰되어 동결 융해후의 난자에서 염색체와 방추사의 이상 빈도가 증가됨이 관찰되었다.찰되었다. 배아이식후 대조군과 실험군에서 각각 2 마리가 임신이되어 정상적인 산자를 분만하였다. 따라서 항동해제에 taxol의 첨가는 동결 융해후의 난자의 배발달율을 증진시킬 수 있었다..8%로 나타나 난할율 및 배반포 발생율에 있어서 융합조건에 따라 큰 차이는 없었으나 1.9㎸/cm, 30$\mu\textrm{s}$ 2회의 조건이 다른 조건들에 비하여 유의적으로 낮았다. 따라서, 체세포와 수핵란 세포질간의 융합율과 배반포 발생에 미치는 영향은 전압보다는 시간에 더 크게 받음을 알 수 있었으며, 이와 같은 결과에서 융합시 시간을 오래 주는 것보다 전압을 높이는 것이 수핵난자의 세포질에 상해를 줄이고 이후 배반포 발생에 유리할 것으로 사료되었다.면에서도 더욱 더 활발할 것으로 기대된다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며

• Asian Journal of Turfgrass Science
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• v.25 no.2
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• pp.133-137
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• 2011

Winterkill can be defined as any injury including freeze stress kill, winter desiccation, and low temperature disease to turfgrass plants that occurs during the winter period. The major damages from winterkill were low temperature kill, crown hydration, and winter desiccation. Low temperature kill is caused by air and soil temperature. Soil temperature affect more severe to turfgrass than air temperature because low soil temperature cause fetal damage to turfgrass crown. Crown hydration is a form of winter injury in which intercellular water within the plant freezes and causes physical injury to the cell membrane and wall. This is eventually resulted in dehydration of cell. Winter desiccation is the death of leaves or whole plants due to drought during the winter period. To reduce winterkill damage, cultivar selection is very important. If changing cultivar is not allowed, cold temperature hardiness needs to be increased by providing nutrients especially phosphorus and potassium in the late fall. Turf cover is effective way to reduce winterkill damage. Remaining snow is positive process to reduce winterkill damage by insulating soil temperature. The previous researches reported many materials as turf cover such as straw, polypropylene, polyester, and wood mat. Aeration and topdressing is one of the process against winterkill. Both methods are mainly conducted to reduce thickness of thatch layer. In recent, relatively new materials called black or winter topdressing sand are used to protect soil temperature from low air temperature and thaw ice crystal that may remain in soil.

• Journal of Chest Surgery
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• v.36 no.4
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• pp.219-228
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• 2003

Background:Liquid nitrogen freezing techniques have already met with widespread success in biology and medicine as a means of long-term storage for cells and tissues. The use of cryoprotectants such as glycerol and dimethylsulphoxide to prevent ice crystal formation, with carefully controlled rates of freezing and thawing, allows both structure and viability to be retained almost indefinitely. Cryopreservation of various tissues has various con-trolled rates of freezing. Material and Method: To find the optimal freezing curve and the chamber temperature, we approached the thermodynamic calculation of tissues in two ways. One is the direct calculation method. We should know the thermophysical characteristics of all components, latent heat of fusion, area, density and volume, etc. This kind of calculation is so sophisticated and some variables may not be determined. The other is the indirect calculation method. We performed the tissue freezing with already used freezing curve and we observed the actual freezing curve of that tissue. And we modified the freezing curve with several steps of calculation, polynomial regression analysis, time constant calculation, thermal response calculation and inverse calculation of chamber temperature. Result: We applied that freezing program on mesenchymal stem cell, chondrocyte, and osteoblast. The tissue temperature decreased according to the ideal freezing curve without temperature rising. We did not find any differences in survival. The reason is postulated to be that freezing material is too small and contains cellular components. We expect the significant difference in cellular viability if the freezing curve is applied on a large scale of tissues. Conclusion: This program would be helpful in finding the chamber temperature for the ideal freezing curie easily.

• Korean Journal of Food Science and Technology
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• v.31 no.3
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• pp.682-687
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• 1999

The destruction of tissues by volume increase at food freezing is accepted as one of the factor responsible for quality damage. For this reason, the internal pressure developed in meats were investigated with a pressure transducer during freezing, frozen storage and thawing. Increasement of 6.33% for volume and $942.17\;kg/cm^2$ for density at $-20^{\circ}C$ for beef were shown. In quick and slow freezing of beef, internal pressure reached to highest point after reached to the lowest point at initial of the zone of ice crystal formation. The internal pressure was approximately $8{\sim}10\;psig$ and pressure difference was about 1 psig, which was bigger in immersion freezing than that of still-air freezing. During frozen storage of pork, internal pressure of $1.84{\sim}2.32\;psig$ occurred repeatedly as a function of sample weight at material temperature difference of ${\pm}1^{\circ}C$. The internal pressure during thawing of pork was decreased slowly after rapid increase to the maximum for less than 5min at the beginning of thawing. Internal pressure value at thawing was higher than that at freezing in most cases. Internal pressure of beef with thermal equalized freezing was about $1{\sim}4\;psig$, which was lower than that of non-thermal equalized freezing. Also, freezing time was shortened to $10{\sim}20%$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.7
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• pp.1055-1061
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• 2014

Innovative freezing technology is currently applied to preserve foodstuffs for long-term storage. Generally, the quality of frozen food is closely related to the types of freezing and thawing processes. In this study, we characterized the physicochemical properties of onions depending on freezing rate. When onions were frozen at $-40^{\circ}C$, freezing rates were 0.1, 0.5, and $0.7^{\circ}C/min$ depending on air-blast quick freezer mode. Onions were thawed by microwave irradiation at 400 W. Hardness of onion dramatically decreased after freezing and thawing compared with blanched onion. However, the fastest freezing rate did not affect hardness. Thawing loss of onion decreased with a faster freezing rate. For morphological observation, onion frozen at a faster rate showed a smaller ice-crystal size. Vitamin C content decreased upon blanching or freezing, but there was no significant difference according to freezing rate. Although free sugar content also decreased upon blanching and freezing, its highest content was at $0.7^{\circ}C/min$ freezing. Among organic acids, malic acid content was highest at $0.7^{\circ}C/min$ freezing. Based on this study, it could be suggested that a faster freezing rate is effective to improve frozen food quality in accordance with preventing tissue damage or minimizing destruction of nutrients.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.4
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• pp.463-470
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• 1998

To improve muscle quality and prolong freshness of sashimi, the effects of freezing-thawing condition on physicochemical and rheological properties of plaice muscle were investigated. Muscle tested were frozen with quick freezing (liquid nitrogen gas) or slow freezing ($-15^{\circ}C$ air). Transition time of zone of ice crystal formation was within 10 minute for quick freezing and 110 minute for slow freezing. Time required for thawing to $0^{\circ}C$ in muscle temperature by various thawing methods was shortest with $25^{\circ}C$ tap water, followed by $15^{\circ}C$ tap water, $10^{\circ}C$ tap water, $25^{\circ}C$ air, $5^{\circ}C$ tap water and $0^{\circ}C$ cold water. Breaking strength of muscle was higher in quickly frozen sample than in slowly frozen sample. According to sashimi term, changes in breaking strength of muscle did not show any difference in quickly frozen sample, while showed significant difference in slowly frozen sample. The remaining content of ATP was not effected by freezing speed, and ATP content was apt to higher in quickly thawed sample than in slowly thawed sample. IMP was the majority of ATP and it's related compounds of sample after freezing and thawing. Collagen matrix was weakened markedly in slowly frozen sample than in quickly frozen sample.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.10
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• pp.1492-1503
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• 2015

This study evaluated the combination effect of various freezing and thawing techniques on the quality and nutritional aspects of onions. Onions were frozen by natural air convection freezing (NCF), air blast freezing (ABF), and liquid nitrogen freezing (LNF). Onions were frozen for 76 min by NCF, 9 min by ABF, and 9 min by LNF. The freezing treatment was stopped when the core temperature reached $-12^{\circ}C$ for NCF and ABF, and $-120^{\circ}C$ for LNF. Frozen samples were thawed through natural air convection thawing, running water thawing, sonication thawing (ST), or microwave thawing. The quality and nutritional aspects of frozen-thawed onions were evaluated by measuring thawing loss, pH, texture, water content, color, and SEM image. ST was found to cause the least loss in onion sample among the tested thawing methods, whereas the freezing methods did not cause any significant loss. In our experiment, thawing is found to be a more critical technique when compared to that of freezing. There were no clear quantifications or trends of pH and water content among different freezing and thawing techniques. The highest total color difference (${\Delta}E$) was observed in the NCF sample. For morphological observation, ABF gave the smallest ice crystal size, as well as minimum cell collapse. Loss of vitamin C, free sugar, and organic acid content was lower in the ABF and ST sample, when compared to other trials. In our study, we found that combination of ABF and ST could preserve the quality and nutritional aspects of frozen-thawed onions better than other methods.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.4
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• pp.307-319
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• 2010

Objective: Vitrification requires a high concentration of cyroprotectant (CPA) and an elevated cooling speed to avoid ice crystal formation. We have evaluated the effect of different combinations of cooling rate and CPA on embryonic integrity (developmental competence) in order to increase the efficiency of vitrification without impairing embryo viabilit. We hypothesized that the combination of CPA or the increase of cooling rates can reduce the concentration of toxic CPA for vitrification. As consequently, we performed experiments to evaluate the effect of various composition of CPA or slush nitrogen ($SN_2$) on the mouse embryonic development following vitrification using low CPA concentration. Methods: Vitrification of mouse embryos was performed with EM grid using liquid nitrogen ($LN_2$) or $SN_2$ and different composition of CPAs, ethylene glycol (EG) and dimethylsulfoxide (DMSO). After vitrification-warming process, their survival and blastocyst formation rates were examined. For analyzing long-term effect, these blastocysts were transferred into the uterus of foster mothers. Results: Survival and blastocyst formation rates of vitrified embryos were higher in EG+DMSO group than those in EG only. Furthermor, the group using $SN_2$ with a lower CPA concentration showed a higher survival of embryos and developmental rates than group using $LN_2$. Conclusion: The combination of EG and DMSO as CPAs may enhance the survival of mouse embryos and further embryonic development after vitrification. $SN_2$ can generate high survival and developmental rate of vitrified/warmed mouse embryos when a lower concentration of CPA was applied. Therefore, these systems may contribute in the improvement of cryopreservation for fertility preservation.

• Food Science and Preservation
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• v.24 no.2
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• pp.168-180
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• 2017

This study was performed to determine the rapid thawing method for reducing the thawing time of frozen pork loins and to examine the effects of supercooling on the microbiological, physicochemical, and sensory qualities of fresh and frozen-thawed pork during storage at -1.5, 4, and $15^{\circ}C$. Forced-air thawing at $4^{\circ}C$ was the most time-consuming process, whereas radio frequency thawing time was the shortest by dielectric heating. The supercooling storage temperature was chosen to be $-1.5^{\circ}C$ because microstructural damages were not observed in the pork sample after cooling at $-1.5^{\circ}C$ for 24 h. Fresh or frozen-thawed pork loins stored at $-1.5^{\circ}C$ had lower drip loss and total volatile base nitrogen, thiobarbituric acid-reactive substance, and Hunter b* levels than loins stored at 4 and $15^{\circ}C$. In addition, the least degree of increase in preexisting microorganisms counts of the fresh or frozen-thawed pork loin samples was obtained during supercooled storage at $-1.5^{\circ}C$. Sensory quality results of fresh and frozen-thawed pork loin samples stored at $-1.5^{\circ}C$ showed higher scores than the samples stored at 4 and $15^{\circ}C$. These data indicate that supercooling at $-1.5^{\circ}C$ in the meat processing industry would be effective for maintaining the quality of pork meats without ice crystal nucleation and formation.