• Clinical and Experimental Reproductive Medicine
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• 제17권1호
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• pp.81-85
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• 1990

We cryopreserved mouse morulae by a simple ultra-rapid method of freezing embryos directly in $LN_2$ after holding 2min in a $LN_2$ vapor, and thawed them in $37^{\circ}C$ water bath. The time requirements for permeation and dehydration by 2.0 M glycerol and 0.2 M sucrose before freezing were studied. When the embryos were equilibrated for 10 min, the optimun post-thaw survival was obtained. Embryos those developed normally to blastocyst after in vitro culture for over 24hrs were regarded as survival ones. Two experiments to assess post-thaw survival following predehydration in various mixtures of glycerol and sucrose were also accomplished. When sucrose was held constant (0.2 M) and glycerol concentration varied (1.5-3.5 M), post-thaw survival was best (78.0%) in 3.0 M glycerol. When glycerol was held constant (3.0M) and sucrose concentration varied (0.0-1.0M), optimun post-thaw survival (78.0%) was found in 0.2 M sucrose.

• 한국도로학회논문집
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• 제15권3호
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• pp.53-63
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• 2013

PURPOSES : This study is to evaluate moisture susceptibility of asphalt mixtures by using non-destructive impact wave and to determine durability so as to decrease the gap between before and after freezing in the future. METHODS : Using non-destructive impact wave, this study is to determine the dynamic modulus of asphalt specimen. Furthermore, the results obtained from two experiment accelerometers are used for the dynamic modulus determination. The dynamic moduli of specimens are compared with those of the freezing-thawing specimens. RESULTS : Test results showed that the dynamic modulus before freezing and thawing environment loads at each temperature dropped about 3.7% after the environmental loads. Furthermore, correlation analysis indicates that transition of dynamic modulus at each point is about 89.59%. CONCLUSIONS: Evaluation of asphalt mixtures using non-destructive impact wave has excellent repeatability and simple equipment for the test. Consequently, the method in the study will be useful for evaluating the characteristics of a various asphalt mixtures.

• 한국수정란이식학회지
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• 제10권3호
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• pp.193-202
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• 1995

In order to improve the cryopreservatory techniques of livestock embryos, the quick freezing method which is directly plunged in liquid nitrogen via prefreezing procedure without freezing machine was carried out for mouse embryos treated with permeable and nonpermeable cryoprotectants. The viability of frozen-thawed embryos were evaluated by FDA vital dye test. The results obtained was summaried as follows: 1. A total of 720 embryos were recovered from frozen embryos for viability test. Evalution of the fluorescein diacetate(FDA) vital dye test with mice embryos were resulted of 2.3 total mean score - evaluted in orderly higher mean grade of P3 453 (63%), P2 133(18%), P1 51(7%) and P0 83(12%). 2. An all-round evalution of these combination, the highest viability was showed in 3M ethylene glycol + 0. 25M trehalose treated with the copper prefreezing. 3. Effects of permeable and nonpermeable cryoprotectants combination were evaluated by means FDA score. 3M ethylene glycol + 0.25M trehalose showed the highest survival rates of 2.8 mean FDA score. 4. Effects of permeable cryoprotectants were evaluated by mean FDA score but the results were not significantly different each other. 5. In evalution of the nonpermeable cryoprotectants, 0. 25M trehalose obtalned higher mean FDA score than of 0.25M sucrose and it was significantly different(P<0.05). 6. There was no significantly difference between copper and stainless-steel in prefreezing procedures.

• 한국임상수의학회지
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• 제4권2호
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• pp.449-455
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• 1987

Quick freezing of bovine embryos was attempted after they were predehydrated at room temperature. Combined solutions of 2M glycerol or 2M ethylene glycol in the presence of either 0.5 or 1.0M sucrose in phosphated buffered saline+20% calf serum were compared. The quick freezing method in which embryos were directly transferred in liquid nitrogen vapor for 2 minutes at - l70$^{\circ}C$ before being plunged into liquid nitrogen was used. Post-thaw survival rates in 2M glycerol and 2M ethylene glycol were high with 0.5 M (55.6% and 53.3%) versus 1.0M(38.1% and 31.6%) sucrose(P < 0.05). But survival rates with 2M glycerol and 2M ethylene glycol were not significantly different. Transfer thawed embryos frozen with 2M glycerol and 2M ethylene glycol by 0.5M sucrose resulted in birthrates of 40.9% and 40.0%, respectively compared to 26.3% and 27.2%, respectively, for 1.0 M sucrose(p<0.05). This was 56.0% for fresh control.

• Journal of Advanced Marine Engineering and Technology
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• 제31권5호
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• pp.622-629
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• 2007

Some worker is occupationally exposed to cold and freezing environment. The cold stimuli in the working environment impose physiological and psychological loads on workers to decrease the task performance. The purpose of this study is to investigate the cold stimuli of cold and freezing stores widely used in Busan can make an effect on human's physiological responses and task performance, experimentally and analytically. In the experiment, 5 workers are selected as subjects, and then their skin temperatures of hand and ear, heart rates, blood pressure, and ring test performances in cold($3^{\circ}C$) and freezing($-22^{\circ}C$) stores were measured for 21 minutes and analyzed by using statistical method. It is observed that a physiological variation and the task performance are significantly influenced by an exposure time as well as a strength of cold stimuli. Also, it is suggested the exposure limiting times for the useful manual work and the performance predict model of the ring tasks. The result of this study will be useful for a fundamental data of which design the standard task time of manual tasks and solve the job placement problem of worker selection and placement in cold environment.

• 한국수산과학회지
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• 제20권3호
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• pp.185-190
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• 1987

부분동결의 유효성을 미생물학적인 측면에서 해명하고자 저온에서 부패의 주역을 담당하고 있는 몇종의 저온세균을 공시균으로 하여서 고등어육액즙에 대한 부패활성을 동일액상이면서 빙점부근에서의 근소한 온도차($0^{\circ}C$와 $-3^{\circ}C$ 과냉각) 및 $-3^{\circ}C$동일 온도에서 상변화(액상-고상)를 중심으로 균의 증식, 사멸, 부패활성 및 K값의 변화를 검토한 결과는 다음과 같다. (1) $25^{\circ}C$ 및 $0^{\circ}C$ 저장에서는 공시균 전부 강한 증식력 및 부패활성을 나타내었으나, 동일액상이면서 $0^{\circ}C$와 근소한 온도차인 $-3^{\circ}C$ 과냉각에서는 공시균 중 Pseudomonas III/IV-NH형의 증식력 및 부패활성이 현저히 억제되었으며, Moraxella 및 F/C도 실험기간을 통하여 점차 감소하는 경향을 나타내었고, 부패활성은 전혀 관찰되지 않았다. (2) $-3^{\circ}C$ 과냉각(액상)과 동일온도이면서 고상인 $-3^{\circ}C$ 동결에서는 전공시균의 증식 및 부패활성이 전혀 관찰되지 않았으며, 저장기간을 통하여 생균수가 감소하는 경향을 나타내었다. (3) 동일액상이면서 $0^{\circ}C$와 근소한 온도차인 $-3^{\circ}C$ 과냉각에서 K값의 증가가 억제되었으며, 더욱 $-3^{\circ}C$의 동온이상인 $-3^{\circ}C$동결(고상)에서는 상의 변화로 인하여 K값의 증가가 현저히 억제되었다. 이상의 결과로부터, $0^{\circ}C$와의 근소한 온도차 및 액상에서 친상으로의 상의 변화에 의한 효소황성의 저하(K값) 및 저온세균의 증식과 부패활성의 억제가 부분동결의 유효성의 큰 요인이 되리라 추측된다.

• 한국수정란이식학회지
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• 제30권3호
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• pp.207-211
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• 2015

미세주입법에 의한 형질전환 마우스 제작에는 대량의 전핵기란을 필요로 한다. 본 실험에서는 심플한 형질전환 마우스 제작방법을 확립하기 위해 초급속 동결한 전핵기란을 공시했다. 초급속 동결법으로 동결한 전핵기 체외수정란 139개를 융해 후 공시하고, ${\beta}-actin/luc^+$ 융합유전자를 미세주입하였다. 주입 조작 후, 형태학적으로 정상적인 수정란 101개(72.6%)를 5마리의 수란암컷 마우스에 이식하였다. 그 결과, 이식한 모든 마우스가 임신하고, 최종적으로 15마리(14.8%)의 산자가 태어났다. 한편, 450개의 체외수정란에 대해 동일한 배아조작 후에 338개(75.1%)가 생존하고 14마리의 수란암컷 마우스에 이식 하였다. 그 중에 78%의 수정암컷 마우스가 임신하고, 54마리(19.1%)의 산자가 태어났다. 태어난 산자에 대해서는 southern blot 법에 의해 염색체 내의 도입유전자의 도입을 확인한 결과, 동결수정란 처리구와 체외수정란구에서 각각 6.6%(1/15), 5.5% (3/54)의 마우스에서 도입유전자의 도입이 확인되었다. 더욱이 두 처리구 전부의 형질전환 마우스의 미부조직에서 도입유전자인 루시페라제 유전자의 발현이 관찰되었다. 이상의 결과에 의해 체외수정란 초급속 동결보존법을 사용한 Tg 마우스 제작방법의 확립을 확인하였고, 이러한 결과들로부터 실험기간의 단축과 작업의 간소화에 크게 이바지할 것으로 사료된다.

• 한국식품과학회지
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• 제29권2호
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• pp.326-336
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• 1997

INP를 포함한 빙핵활성세균을 식품 가공 단계에 직접 이용하기 위해 빙핵활성을 최대한 유지시키면서 완전 살균하는 방법을 찾기 위해 가열, 고압, 방사선으로 처리한 후 생균수 및 빙핵활성을 측정하였다. 균주는 Pseudomonas syringae, Xanthomonas campestris와 INP유전자를 포함한 plasmid가 재조합된 Escherichia coli JM109/pEIN229, Gluconobacter oxydans/pKIN230를 사용하였다. 빙핵활성을 T90 (drop의 90%가 어는 온도)으로 나타내었을 때 방사선 조사시 $0.3^{\circ}C{\sim}0.7^{\circ}C$, 초고압 처리시 $1^{\circ}C{\sim}2^{\circ}C$, 가열 처리시 $4^{\circ}C{\sim}7^{\circ}C$가 감소하여 방사선 조사에 의한 살균시 빙핵활성의 감소가 가장 적은 것으로 나타났다. 각 살균점에서의 cumulative ice nucleation activity spectra를 측정한 결과 A부류의 빙핵($-5^{\circ}C$ 이상에서 빙핵으로 작용)의 수는 방사선처리시 거의 감소가 없었으며 초고압처리시 90% 이상 감소하였으며 가열처리시 존재하지 않았다. 증류수에 방사선조사로 살균된 빙핵활성세균을 첨가한 후 DSC를 이용해 thermogram을 얻었다. 빙핵활성세균을 첨가하지 않은 증류수보다 $11^{\circ}C{\sim}15^{\circ}C$ 정도 높은 온도에서 발열 피크가 형성되기 시작했으나 흡열 피크는 큰 차이를 보이지 않았으며, 발열량 및 흡열량은 감소했다. 한편, DSC를 이용해 측정된 빙핵활성은 drop freezing method에 의해 측정된 값과 높은 연관성$(R^{2}>0.993,\;p<0.0001)$을 보여 간편하고 정확한 빙핵활성 측정법으로의 사용 가능성을 제시했다.

• 한국식품과학회지
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• 제6권2호
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• pp.70-74
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• 1974

한우(韓牛)의 round muscle과 오징어 근육(筋肉)을 여러 가지 조리조건(調理條件)으로 처리(處理)하여 pepsin에 의한 in vitro 소화율(消化率)과 소화과정중(消化過程中)의 amino태(態) N, 유기용매(有機溶媒) 처리(處理)가 소화율(消化率)에 미치는 영향(影響)을 AOAC에 의한 semi-micro Kjeldahl법(法)으로 실험(實驗)한 결과(結果)는 다음과 같다. 1. 소화율(消化率)은 우육(牛肉)에서 autoclaving, frying, raw, freezing, roasting, boiling, dry heating의 순서이며 오징어육(肉)에서는 raw, autoclaving, boiling, freezing, frying, dry heating, roasting의 순서로 높았다. 2. Amino태(態) N는 우육(牛肉), 오징어육(肉) 공(共)히 소화율(消化率)이 높은 순서로 비례해서 증가(增加)한다. 3. 우육(牛肉)과 오징어육(肉)의 raw condition에서 ether처리(處理)가 소화율(消化率)에 거의 영향을 미치지 않는다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.265-271
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• 2011

This study was undertaken to find out the effect of methyl-beta-cyclodextrin (MBCD) on cold shock and membrane cholesterol quantity of sperm during the freezing process in miniature pigs. For this study, semen ejaculated from PWG M-type miniature pig was diluted that freezing solution (with egg yolk group) and m-Modena B (without egg yolk group) treated with 0, 1, 5, 10 and 20 mM MBCD before freezing process. The diluted semen was monitored sperm ability at room temperature, after cooled until $5^{\circ}C$ and after forzen-thawed for cold shock test of spermatozoa. Also, membrane cholesterol of sperm was extracted by folch solution at the same time sperm ability was assessed for viability and acrosomal status. The membrane cholesterol quantity was measured by thin-layer chromatography (TLC) method. The result, viability and acrosome integrity in semen diluted without egg yolk groups were decreased at all temperature range by increasing of MBCD concentration. In particular, sperm of egg yolk group was showed that significantly higher viability and lower acrosome damage when treated with 5 mM MBCD (p<0.05). The results of TLC experiment, cholesterol amounts were increased with MBCD cocentration in egg yolk, and decreased with MBCD concentration in m-Modena B. In cryopreservation efficiency, there was no significant difference at viability, and acrosomal state of sperm in 5 mM MBCD concentration was significantly lower in acrosome damage than other groups (p<0.05). Therefore, the addition MBCD in egg yolk was protected spermatozoa from cold shock injury. This protective effect of MBCD may be due to addition of sperm membrane cholesterol.