• Title/Summary/Keyword: thawing after freezing

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Protease Activities in Tenderizing Effect of Vegetables used as Cooking Material (조리용 채소의 단백분해효소 활성 및 연육효과)

  • Suh, Hyung-Joo;Chung, Soo-Hyun;Choi, Yang-Mun;Cho, Won-Dai
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.883-887
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    • 1998
  • Protease activities were measured in vegetables used as cooking material or a side dish. Proteases of green pepper (Kwari and Chungyang), perilla leaves, soybean sprout and mungbean sprout were showed high activities. Especially, protease in soybean sprout was the highest activity among them. After dialysis, remaining activities in mungbean sprout, green pepper (Kwari and Chungyang) and perilla leaves were 12, 23, 45% and 37%. In the results of thawing after freezing the proteases, remaining activities in sprout of mungbean and soybean were 100% and 65%. Protease in soybean sprout was showed higher activity and stability than others. Proteolytic effects of soybean sprout on myofibrillar and sarcoplasmic protein were showed higher than stroma protein. In SDS-PAGE, myosin heavy chain, actin and tropomyosin were hydrolyzed with increasing time. Protease activity was rapidly increased with increasing meat concentration in a early proteolysis reaction, but was slightly increased in later.

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Genome-wide association study for frozen-thawed sperm motility in stallions across various horse breeds

  • Nikitkina, Elena V.;Dementieva, Natalia V.;Shcherbakov, Yuri S.;Atroshchenko, Mikhail M.;Kudinov, Andrei A.;Samoylov, Oleg I.;Pozovnikova, Marina V.;Dysin, Artem P.;Krutikova, Anna A.;Musidray, Artem A.;Mitrofanova, Olga V.;Plemyashov, Kirill V.;Griffin, Darren K.;Romanov, Michael N.
    • Animal Bioscience
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    • v.35 no.12
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    • pp.1827-1838
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    • 2022
  • Objective: The semen quality of stallions including sperm motility is an important target of selection as it has a high level of individual variability. However, effects of the molecular architecture of the genome on the mechanisms of sperm formation and their preservation after thawing have been poorly investigated. Here, we conducted a genome-wide association study (GWAS) for the sperm motility of cryopreserved semen in stallions of various breeds. Methods: Semen samples were collected from the stallions of 23 horse breeds. The following semen characteristics were examined: progressive motility (PM), progressive motility after freezing (FPM), and the difference between PM and FPM. The respective DNA samples from these stallions were genotyped using Axiom Equine Genotyping Array. Results: We performed a GWAS search for single nucleotide polymorphism (SNP) markers and potential genes related to motility properties of frozen-thawed semen in the stallions of various breeds. As a result of the GWAS analysis, two SNP markers, rs1141327473 and rs1149048772, were identified that were associated with preservation of the frozen-thawed stallion sperm motility, the relevant putative candidate genes being NME/NM23 family member 8 (NME8), olfactory receptor family 2 subfamily AP member 1 (OR2AP1), and olfactory receptor family 6 subfamily C member 4 (OR6C4). Potential implications of effects of these genes on sperm motility are herein discussed. Conclusion: The GWAS results enabled us to localize novel SNPs and candidate genes for sperm motility in stallions. Implications of the study for horse breeding and genetics are a better understanding of genomic regions and candidate genes underlying stallion sperm quality, and improvement in horse reproduction and breeding techniques. The identified markers and genes for sperm cryotolerance and the respective genomic regions are promising candidates for further studying the biological processes in the formation and function of the stallion reproductive system.

Effects of N-Methylacetamide on the Viability, Fertility and Hatchability of Cryopreserved Ogye (Korean Native Black Fowl) Semen (N-Methylacetamide 동결보호제가 오계 동결정액의 생존성, 수정 및 부화율에 미치는 영향)

  • Choi, Jin Seok;Kim, Sung Woo;Shin, Dan-Bi;Ko, Yeoung-Gyu;Do, Yoon-Jung;Kim, Dong-Hun;Kong, Il-Keun;Park, Soo-Bong
    • Korean Journal of Poultry Science
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    • v.39 no.4
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    • pp.291-295
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    • 2012
  • The use of methylacetamide (MA) as a cryoprotective agent for freezing Korean Native Black rooster Ogye semen was examined with artificial insemination. The diluted Ogye semen with HS-1 was subjected for 2 step dilution method of cryopreservation in which the final concentration of MA was adjusted to 7.5%. The sperm viability after thawing was reduced from $95.17{\pm}0.93%$ to $55.93{\pm}1.38%$ which was confirmed by live-death analysis based on Fluorescence-Activated Cell Sorting (FACS). The rates of fertilized eggs with fresh or frozen-thawed semen were reduced from $94.98{\pm}3.93%$ to $66.36{\pm}8.43%$ at day 7 with significant difference. However, the hatching rates of experiments at day 21 did not shown difference between $92.64{\pm}2.33%$ and $90.45{\pm}8.05%$ (P<0.05). With these results, the utilization of MA for freezing of Ogye spermatozoa could affect on viability of frozen-thawed semen but not on the fertility of lain eggs and hatchability of fertilized eggs and also provide possible tools of freezing for poultry genetic resource conservation.

Variationsin Air and Ground Temperatures During a Frozen Season in the Subalpine Zone of Mt. Halla (한라산 아고산대의 동결기 기온 및 지온변화)

  • Kim, Taeho
    • Journal of The Geomorphological Association of Korea
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    • v.20 no.3
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    • pp.95-107
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    • 2013
  • In order to examine the temperature regime responsible for periglacial processes, air and ground temperatures were monitored from October 2010 to May 2011 at a subalpine bare patch (1,710m asl) of Mt. Halla. Four thermistor sensor probes were installed at 55 cm above a ground surface and depths of 2 cm, 10 cm, and 20 cm, respectively. A mean air temperature is $-0.1^{\circ}C$, while mean ground temperatures are $1.8^{\circ}C$ at 2 cm, $2.6^{\circ}C$ at 10 cm and $3.2^{\circ}C$ at 20 cm deep. A mean monthly ground temperature at 2 cm deep demonstrates below $0^{\circ}C$ successively from January to March, while those at 10 cm and 20 cm deep show no sub-zero temperature. A total of 72 freeze-thaw cycle was observed in air temperature. However, the numbers in ground temperature reduced into 17 at 2 cm, 8 at 10 cm, and 3 at 20 cm deep. The cycles of air temperature and ground temperature at 2 cm deep mostly fluctuated diurnally, while those of ground temperature at 10 cm and 20 cm deep exhibited a several-daily oscillation. Snow cover over 55 cm high remained from January to early April, and it seemed to disappear completely on April 16. A seasonal frost of at least 2 cm thick was formed on late December and the isotherm of $0^{\circ}C$ descended slowly into 10 cm deep on late March to early April due to the insulating snow cover. It showed the maximum freezing depth of 20 cm on April 7 to 14 and then thawed rapidly so that the frozen ground did not longer after April 17. Periglacial processes are predominant during a freezing period than a thawing period when the ground surface is still covered with snow. The periglacial mass movement in the subalpine zone of Mt. Halla is mainly generated by frost creep in terms of the occurrence depth of diurnal freeze-thaw cycle and the maximum freezing depth of ground.

Dog Sperm Cryopreservation Using Glucose in Glycerol-free TRIS: Glucose Concentration, Exposure Time (Glycerol-free TRIS 배지내 glucose를 이용한 개 정자 동결: 포도당 농도, 노출시간)

  • Yu, Il-Jeoung
    • Journal of Veterinary Clinics
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    • v.30 no.6
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    • pp.442-448
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    • 2013
  • The aim of the present study was to develop glycerol-free TRIS extender using glucose for dog sperm cryopreservation. We determined the appropriate concentration of glucose in glycerol-free TRIS and the exposure time in glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$. Ejaculates of six dog sperm were cooled in glycerol-free TRIS through $4^{\circ}C$ for 100 min, cooled at $4^{\circ}C$ in TRIS with different glucose concentrations 0 M, 0.04 M, 0.1 M, 0.2 M and 0.3 M, respectively for 30 min followed by cryopreservation. After thawing at $37^{\circ}C$ for 25 sec, membrane and acrosome integrities of dog sperm were evaluated. In addition, the effect of exposure time (10, 30, 50 and 70 min) of sperm to glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$ on progressive motility, viability, and DNA integrity following sperm cryopreservation was studied. Membrane integrity and acrosome integrity were assessed by 6-carboxyfluoresceindiacetate (6-CFDA)/propidium iodide (PI) fluorescent staining and Pisum sativum agglutinin conjugated to fluorescein isothiocyanate, respectively. DNA integrity was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling, using flow cytometry. Sperm frozen in glycerol-free TRIS supplemented with 0.2 M or 0.3 M glucose have an intact plasma membrane (CFDA+/PI-) after cryopreservation than sperm frozen in the extenders with lower glucose concentrations (p<0.05). Acrosome integrity was significantly higher in the 0.3 M group than less than 0.1 M groups (p<0.05). The sperm DNA fragmentation index did not differ according to exposure time, although progressive motility was significantly higher in the 50 min exposure group than the other groups (p<0.05). These results indicate that cryopreservation of dog sperm is feasible and yields more motile sperm following freezing and thawing in glycerol-free TRIS containing 0.3 M glucose with the exposure time for 50 min at $4^{\circ}C$.

Effects of Mitochondria-targeted Antioxidant MitoTEMPO on the Kinetic Characteristics of Frozen-Thawed Boar Sperm (동결-융해 정자의 운동학적 특성에 대한 MitoTEMPO의 영향)

  • Cho, Eun Seok;Kim, Jeong A;Jeong, Yong Dae;Choi, Yo Han;Hong, Jun Ki;Kim, Young Sin;Chung, Hak Jae;Baek, Sun Young;Sa, Soo Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.3
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    • pp.199-205
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    • 2020
  • Cryopreservation of semen is useful for animal breeding via artificial insemination (AI). However, the use of frozen-thawed boar semen is limited due to cryodamage. The aim of this study was to investigate the effects of different concentrations of MitoTEMPO (a mitochondria-targeted antioxidant) in lactose-egg yolk (LEY) extenders on kinetic characteristics of frozen-thawed boar sperms. Semen samples were collected from mature Duroc boars (2~3 years old) and cryopreserved in LEY extenders containing 0, 0.5, 5, 50, and 500 μM MitoTEMPO. The kinetic characteristics of frozen-thawed sperms were determined 0 and 30 min after thawing using computer-assisted sperm analysis (CASA). Results indicated that sperm motility immediately after thawing was significantly higher with 5 and 50 μM (50.46±2.71% and 46.96±2.66%, respectively) than with 500 μM MitoTEMPO (35.40±2.95%) (P<0.05). However, there were no significant differences in other kinetic characteristics except motility. In conclusion, the addition of MitoTEMPO to the sperm freezing extender may have a beneficial effect on motility of post-thawed boar semen.

Establishment of Bovine Ovum Bank : I. Full Term Development of Vitrified In Vitro Matured Hanwoo (Korean Cattle) Oocytes by Minimum Volume Cooling (UC) Method (소 난자 은행 설립 : I. MVC 방법으로 초자화 동결된 한우 미성숙 난자의 개체 발생능 조사)

  • Kim, E.Y.;Kim, D.I.;Rhee, M.G.;Weon, Y.S.;Nam, H.K.;Lee, K.S.;Park, S.Y.;Park, E.M.;Yoon, J.Y.;Heo, Y.T.;Cho, H.J.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.1-7
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    • 2001
  • This study was to test whether in vitro matured Hanwoo oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5~10 min, exposed in EG30 for 30 sec, each oocyte was individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures 〔1.0 M sucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS〕 at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were pregnant and three of them were ongoing-pregnant by manual palpation at 250 days after transfer. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.

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Development of Deterioration Prediction Model and Reliability Model for the Cyclic Freeze-Thaw of Concrete Structures (콘크리트구조물의 반복적 동결융해에 대한 수치 해석적 열화 예측 및 신뢰성 모델 개발)

  • Cho, Tae-Jun;Kim, Lee-Hyeon;Cho, Hyo-Nam
    • Journal of the Korea Concrete Institute
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    • v.20 no.1
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    • pp.13-22
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    • 2008
  • The initiation and growth processes of cyclic ice body in porous systems are affected by the thermo-physical and mass transport properties, as well as gradients of temperature and chemical potentials. Furthermore, the diffusivity of deicing chemicals shows significantly higher value under cyclic freeze-thaw conditions. Consequently, the disintegration of concrete structures is aggravated at marine environments, higher altitudes, and northern areas. However, the properties of cyclic freeze-thaw with crack growth and the deterioration by the accumulated damages are hard to identify in tests. In order to predict the accumulated damages by cyclic freeze-thaw, a regression analysis by the response surface method (RSM) is used. The important parameters for cyclic freeze-thawdeterioration of concrete structures, such as water to cement ratio, entrained air pores, and the number of cycles of freezing and thawing, are used to compose the limit state function. The regression equation fitted to the important deterioration criteria, such as accumulated plastic deformation, relative dynamic modulus, or equivalent plastic deformations, were used as the probabilistic evaluations of performance for the degraded structural resistance. The predicted results of relative dynamic modulus and residual strains after 300 cycles of freeze-thaw show very good agreements with the experimental results. The RSM result can be used to predict the probability of occurrence for designer specified critical values. Therefore, it is possible to evaluate the life cycle management of concrete structures considering the accumulated damages due to the cyclic freeze-thaw using the proposed prediction method.

Evaluation of Environment Friendly High Performance Ternary Cement Concrete Deck Overlay Pavement by Experimental Construction (시험시공을 통한 친환경 고성능 3성분계 시멘트 콘크리트 교면 포장의 성능 평가)

  • Choi, In-Hyeok;Kim, Dae-Seong;Lee, Jun-Ho
    • International Journal of Highway Engineering
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    • v.13 no.2
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    • pp.85-93
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    • 2011
  • This study experimented to evaluate the environment friendly high performance ternary cement concrete deck overlay pavement using mineral admixture such as fly ash and ground granulated blast-furnace slag. It was measured to find best binder mixing according to replacement rate of mineral admixture with compressive strength and flexural strength. After finding best binder, it is also experimented to evaluate durability on chloride penetration resistance, freezing- thawing resistance, scaling resistance of deicing chemicals, abrasion resistance, alkali-silica reactivity test and bonded environment friendly high performance ternary cement concrete deck overlay pavement experimented to evaluate bonded old deck and new concrete overlay pavement using special polymer cement mortar. In additions, bonded environment friendly high performance ternary cement concrete deck overlay pavement by experimental construction was evaluated at interchange bridge of North Yeoju. Result, examination was indicated better binding with binder replacement of cement 70%, ground granulated blast-furnace slag 15% and fly ash 15%. And special polymer cement mortar used in old deck and new overlay concrete was indicated better bonding both laboratory and construction.

An Experimental Study on Properties of Concrete Using Latent Heat Binder (잠열성 결합재를 활용한 콘크리트의 특성에 관한 실험적 연구)

  • Kim, Yong-Ro;Kim, Do-Su;Khil, Bae-Su;Kim, Ook-Jong;Lee, Do-Bum
    • Journal of the Korea Concrete Institute
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    • v.20 no.5
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    • pp.661-668
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    • 2008
  • It is necessary to develop a new technology for effectively reducing hydration heat and controlling thermal cracking caused increasing construction of large size massive concrete structures such as mat foundation of high-rise building, grandiose bridge, and LNG tank. Therefor, to develop a new technology for reducing hydration heat of large size massive concrete in this study, after developing the latent heat binder for controling hydration heat of concrete by application of latent heat material, it was investigated basic properties and durability such as slump, air content and compressive strength, shrinkage properties, permeability, freezing and thawing resistance, corrosion, and hydration heat generation properties of concrete using latent heat binder. As a test result, it was confirmed that latent heat binder was not affected adversely the basic property and durability of concrete, and was advanced on the reduction of hydration heat and control of thermal crack. It is expected to be applied as the excellent technology on the management of hydration heat and thermal crack in large size mass concrete structures.