• Clinical and Experimental Reproductive Medicine
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• 제46권2호
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• pp.67-75
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• 2019

Objective: Sperm cryopreservation has been widely used in assisted reproductive technology, as it offers great potential for the treatment of some types of male infertility. However, cryopreservation may result in changes in membrane lipid composition and acrosome status, as well as reductions in sperm motility and viability. This study aimed to evaluate sperm DNA fragmentation damage caused by conventional freezing using the sperm chromatin dispersion test. Methods: In total, 120 fresh human semen samples were frozen by conventional methods, using SpermFreeze Solution as a cryoprotectant. Routine semen analysis and a Halosperm test (using the Halosperm kit) were performed on each sample before freezing and after thawing. Semen parameters and sperm DNA fragmentation were compared between these groups. Results: There was a significant decrease in sperm progressive motility, viability, and normal morphology after conventional freezing (32.78%, 79.58%, and 3.87% vs. 16%, 55.99%, and 2.55%, respectively). The sperm head, midpiece, and tail defect rate increased slightly after freezing. Furthermore, the DNA fragmentation index (DFI) was significantly higher after thawing than before freezing (19.21% prior to freezing vs. 22.23% after thawing). Significant increases in the DFI after cryopreservation were observed in samples with both normal and abnormal motility and morphology, as well as in those with normal viability. Conclusion: Conventional freezing seems to damage some sperm parameters, in particular causing a reduction in sperm DNA integrity.

• Asian-Australasian Journal of Animal Sciences
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• 제22권3호
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• pp.343-349
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• 2009

This study was designed to simplify a cryopreservation program for embryonic stem cells (ESCs) by selection of cooling method and cryoprotectant. Commercially available mouse E14 embryonic stem cells (ESCs) were cryopreserved with various protocols, and morphology and viability of the frozen-thawed ESCs and their reactive oxygen species (ROS) production were subsequently monitored. Post-thaw colony-formation of ESCs was detected only after a slow freezing using dimethyl sulfoxide (DMSO) by stepwise placement of a freezing container into a $-80^{\circ}C$ deep freezer and subsequently into -$196^{\circ}C$ liquid nitrogen, while no proliferation was detected after vitrification. When the simplified protocol was employed, the replacement of DMSO with a mixture of DMSO and ethylene glycol (EG) further improved the post-thaw survival. ROS generation in ESCs frozen-thawed with the optimized protocol was not increased compared with non-frozen ESCs. The use of fresh mouse embryonic fibroblasts as feeder cells for post-thaw subculture did not further increase post-thaw cell viability. In conclusion, a simplified slow-freezing program without employing programmable freezer but using DMSO and EG was developed which maintains cell viability and colony-forming activity of ESCs during post-thaw subculture.

• 한국수정란이식학회지
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• 제10권3호
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• pp.203-208
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• 1995

This study was carried out to efficiently use the ultrarapid freezing method in the cryopreservation of mouse ova. For this, the effects of dehydration method, oval vigour and $0^{\circ}C$ controlling method on post-thawing viability were investigated. Fresh mouse ova were dehydrated in mPBS with 3.5M DMSO and /or 0.25M sucrose, and directly immersed in L$N_2$ for ultrarapidly freezing. The frozen ova were thawed at 37$^{\circ}C$, rehydrated in mPBS with 0.25M sucrose, and then repeatedly washed in HAM's Fl0 before evaluating the morphological normality of frozen-thawed ova. The results obtained showed that there was difference between treatments in a experiment. 1) The post-thawing viability of ova dehydrated in multi-step (48.4$\pm$13.8%) was higher than that of ova in two-step (40.9$\pm$14.0%). 2) The post-thawing viability of fertilized ova (87$\pm$14.0%) was significantly(p<0.0l) higher than that of unfertilized ova (5.4$\pm$5.4%). 3) The post-thawing viability of ova dehydrated and rehydrated using a cooling machine (95.8$\pm$4.2%) was significantly(p<0.05) higher than that on ice(84.1$\pm$9.9). In conclusion, in order to efficiently cryopreserve ova in vitro with ultrarapidly freezing method, highly viable embryos should be selected, heavy osmotic shock to the dehydrating ova should be avoided, and embryos in high osmotic condition were dehydrated and rehydrated in a constantly low temperature.

• 한국수산과학회지
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• 제6권3_4호
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• pp.101-111
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• 1973

생 전복을 액체질소동결, 송풍동결, 반송풍동결, 정지공기동결의 네가지 동결방법으로 동결하여 $-20^{\circ}C$에서 3개월간 저장하여 동결방법에 의한 전복근육의 물성적 변화, 단백질의 변성 등을 검토하여 다음과 같은 결과를 얻었다. 1) 생전복의 사후경직은 다른 어류보다 빨랐다. 2) 압측시험치, 전단시험치는 액체질소동결을 제외하고는 동결방법에 의한 차는 없었고, 저장기간이 길어짐에 따라 약간 감소하였다. 3) 염가용성질소는 동결저장기간중 용출량이 완만하게 감소하였다. 4) 유리수의 유출량은 패주와 다리 사이에는 차가 있었고, 저장기간중 패주는 유리수가 증가하는 경향이었다. 5) Texture의 Panel test 결과 저장기간을 통하여 Texture가 감소하는 경향이었으며, Texture의 변화는 유리수의 변화(P<0.01), 단백질의 변성(P<0.05), 압축시험치의 변화(P<0.05)와 유의의 상관관계가 있었다. 본 논문을 지도하여 주신 동경수산대학 교수 전중 화부 박사 그리고 논문의 작성에 조언해 주신 부산 수산대학 식품공학과 이응호 박사님께 감사의 뜻을 표한다.

• 한국콘크리트학회:학술대회논문집
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• 한국콘크리트학회 2004년도 추계 학술발표회 제16권2호
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• pp.221-224
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• 2004

Several factors of concrete durability decline factor are acted as complex deterioration and is happened not that happen by simplicity deterioration. Specially, in case of sea construction, as complex salt damage, carbonation and freezing & thawing, concrete surface and pore structure is deteriorated. Therefore, analyzing concrete carbonation and pore structure after freezing and thawing test by fresh water and sea water in this research, we wish to study about acceleration of decline of durability and complex deterioration by concrete surface deterioration in sea environment.

• 농업과학연구
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• 제33권2호
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• pp.129-140
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• 2006

There are need to develop of merchandise of value added fresh ginseng because of high consciousness level of consumer and enlarge of markets for high quality products. The fresh ginseng after harvest was distributed to farmer partually but in general, it was to market by consigner or wholsaler directly after harvest. There were a high difference on storage period of fresh ginseng in different harvesting seasons. The reduction of value of commodities of fresh ginseng for storage period was caused by decomposition and tender of tissue. The storage temperature was under the freezing point and the packing method was sealing tightly by plastic film. As the quality of fresh ginseng was defined by naked eye, it was difficult to sort the quality of ginseng directly harvest.

• 한국콘크리트학회:학술대회논문집
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• 한국콘크리트학회 2003년도 가을 학술발표회 논문집
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• pp.527-532
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• 2003

When fresh concrete is exposed to sufficiently low temperature, the free water in the concrete is cooled below its freezing point and transforms into ice, which causes decrease in compressive strength of concrete. Of the many influencing factors on the loss of compressive strength, the age of concrete at the beginning of freezing, water-cement ratio, and cement-type are significantly important. The objective of this study is to examine how the these factors affect the compressive strength of concrete frozen at early ages. The results from the tests showed that as age at the beginning of freezing is delayed and water-cement ratio is low, the loss of compressive strength decreases. In addition, concrete made with high-early-strength cement is less susceptible to frost damage than concrete made with ordinary portland cement.

• 한국건축시공학회:학술대회논문집
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• 한국건축시공학회 2013년도 추계 학술논문 발표대회
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• pp.32-33
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• 2013

This study was a result of laboratory test to confirm the porperties of concrete containing the domestic artificial lightweight aggregate. The domestic artificial lightweight aggregate is made with bottom ash which waste material in the thermal power plant. In the experimental result air contents of fresh concrete was measured lower than other artificial lightweight aggregate. This air contents is important for retaining the resistance of freezing and thawing. Therefore air contents of concrete will be considered for retaining the resistance of freezing and thawing when manufacture the concrete containing the domestic artificial aggregate.

• Clinical and Experimental Reproductive Medicine
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• 제49권2호
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• pp.117-126
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• 2022

Objective: This study examined whether the addition of triple antioxidants (3A)-10 µM acetyl-L-carnitine, 10 µM N-acetyl-L-cysteine, and 5 µM α-lipoic acid-in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa. Methods: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated. Results: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively). Conclusion: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa.

• 한국축산식품학회지
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• 제24권2호
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• pp.151-155
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• 2004

본 연구는 미토콘드리아 malate dehydrogenase활성을 이용하여 냉장계육과 냉동계육의 판별법을 개발하기 위하여 실시되었다. 단백질의 함량은 8.5mg/mL에서 12.7mg/mL 범위로 나타났으며, 동일 저장 기간 동안의 온도 별 차이점은 모든 부위에서 1일에서 15일 저장기간이 길어짐에 따라 단백질함량이 늘어나는 경향을 보인다. 냉장육의 보수력은 저장 기간 1일 60.5%에서 15일 33.9%로 감소하였으며 유의적 차이는 없었다(p＞0.05). 냉장계육과 냉동계육은 저장 기간이 길어짐에 따라 육즙손실량이 증가하였으며 유의적 차이는 없었다.(p＞0.05). 냉장계육의 육즙손실량이 냉동계육의 육즙손실량보다 낮게 나타났다. 냉장계육과 냉동계육 간의 mitochondrial malate dehydrogenase의 활성은 냉동계육이 냉장계육보다 효소활성이 높게 나타났다. 따라서 본 연구결과로부터 mitochondrial malate dehydrogenase의 활성 여부로 계육의 냉장, 냉동 유무를 판별하는데 유효한 지표로 사용 가능한 것으로 사료되었다.