• Title/Summary/Keyword: Thawing method

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Development of a new mini straw for cryopreservation of boar semen

  • Almubarak, Areeg;Osman, Rana;Lee, Seongju;Yu, Iljeoung;Jeon, Yubyeol
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.2
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    • pp.113-120
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    • 2022
  • Sperm cryopreservation is a fundamental process for the long-term conservation of livestock genetic resources. Yet, the packaging method has been shown, among other factors, to affect the frozen-thawed (FT) sperm quality. This study aimed to develop a new mini-straw for sperm cryopreservation. In addition, the kinematic patterns, viability, acrosome integrity, and mitochondrial membrane potential (MMP) of boar spermatozoa frozen in the developed 0.25 mL straw, 0.25 mL (minitube, Germany), or 0.5 mL (IMV technologies, France) straws were assessed. Post-thaw kinematic parameters were not different (experiment 1: total motility (33.89%, 32.42%), progressive motility (19.13%, 19.09%), curvilinear velocity (42.32, 42.86), and average path velocity (33.40, 33.62) for minitube and the developed straws, respectively. Further, the viability (38.56%, 34.03%), acrosome integrity (53.38%, 48.88%), MMP (42.32%, 36.71%) of spermatozoa frozen using both straw were not differ statistically (p > 0.05). In experiment two, the quality parameters for semen frozen in the developed straw were compared with the 0.5 mL IMV straw. The total motility (41.26%, 39.1%), progressive motility (24.62%, 23.25%), curvilinear velocity (46.44, 48.25), and average path velocity (37.98, 39.12), respectively, for IMV and the developed straw, did not differ statistically. Additionally, there was no significant difference in the viability (39.60%, 33.17%), acrosome integrity (46.23%, 43.23%), and MMP (39.66, 32.51) for IMV and the developed straw, respectively. These results validate the safety and efficiency of the developed straw and highlight its great potential for clinical application. Moreover, both 0.25 mL and 0.5 mL straws fit the present protocol for cryopreservation of boar spermatozoa.

Properties of Iron Powder and Activated Carbon mixed Matrix for the Improvement of Cold Weather Concrete (한중콘크리트 개선을 위한 철가루와 활성탄 혼입 경화체 기초연구)

  • Kim, Won-Jong;Kim, Won-Sik;Kim, Gyu-Yong;Lee, Sang-Soo
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2022.11a
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    • pp.175-176
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    • 2022
  • By studying the characteristics of matrix insulated through heat generated through oxidation of iron powder, the basic research results on the possibility of buffering and applicability of Cold weather concrete as a curing method are presented. In order to prevent freezing due to a sharp decrease in temperature in the initial stage of curing, iron powder (Fe), powder activated carbon, which is a small amount of porous carbonaceous adsorbent, and salt (NaCl) as an oxidizing agent are replaced with iron powder admixture. As the curing temperature increases, the strength tends to increase, and when replacing the admixture at the same curing temperature, the strength slightly decreases. This is determined as a result of generating iron oxide through an oxidation reaction of iron powder, activated carbon, and NaCl generating a large amount of pores in the matrix. In addition, the internal temperature tends to increase as the mixing substitution rate increases, and it is judged that the oxidation heat of the iron powder mixture affects the increase of the internal temperature during curing. The higher the replacement rate of the iron powder mixture, the slightly lower the strength, but it is determined that freezing and melting that may occur in the early stage of curing can be prevented due to an increase in the initial internal temperature.

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Enhancement of cryopreserved rooster semen and fertility potential after oral administration of Thai ginger (Kaempferia parviflora) extract in Thai native chickens

  • Vibuntita Chankitisakul;Supakorn Authaida;Wuttigrai Boonkum;Sarunya Tuntiyasawasdikul
    • Animal Bioscience
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    • v.37 no.7
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    • pp.1177-1184
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    • 2024
  • Objective: Semen cryopreservation is an effective method of preserving genetic material, particularly in native chicken breeds facing a substantial decline. In this study, we evaluated the quality of frozen/thawed rooster semen treated with different concentrations of oral administrations of black ginger (Kaempferia parviflora: KP) extract and determined its fertility. Methods: Thirty-two Thai native roosters (Pradu Hang Dum, 42 weeks old) were used in this study. The treatments were classified into four groups according to the concentration of KP extract administered to the roosters: 0, 100, 150, and 200 mg/kg body weight. The quality of fresh semen was analyzed before cryopreservation. Post-thaw sperm quality and fertility potential were determined. Also, lipid peroxidation was determined. Results: The results showed that sperm concentration and movement increased in roosters treated with 200 mg/kg of KP extract (p<0.05). The malondialdehyde (MDA) in the roosters receiving 200 mg/kg KP extract was lower than that in the other but had an insignificant difference within the KP treatment groups (p>0.05). The highest MDA levels were observed in the control group (p<0.05). The percentage of motile sperm (total motility and progressive motility) after semen thawing was higher in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). MDA levels decreased significantly in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). Fertility and hatchability were greater in the KP150 and KP200 groups than in the KP100 and control groups (p<0.05). Conclusion: The optimal amount of KP extract influencing initial sperm quality was determined to be 200 mg/kg. However, 150 mg/kg was the optimal low dosage of KP extract administration that maintained sperm quality and fertility following semen cryopreservation.

Effect of seeding on post-thaw motility and viability of canine frozen sperm (견정액 동결시 seeding처리가 융해후 정자의 활력 및 생존률에 미치는 효과)

  • 김종호;이필돈;유일정;김용준
    • Korean Journal of Veterinary Service
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    • v.18 no.3
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    • pp.1-12
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    • 1995
  • To investigate effect of seeding on post-thaw motility and viability of canine spermatozoa, the semen from male dogs which had been proved to be fertile in the past were frozen and seeded during freezing process. Post-thaw motility and viability of canine sperm which were frozen and seeded were investigated according to different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$, or $-l5^{\circ}C$ and also according to different concentration of glycerol of 2%, 5% and 10%. In addition, post-thaw motility of canine sperm frozen by direct freezing in a deep freezer or programmed freezing in a programmed cell freezer was investigated. Post-thaw motility of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}C$ showed considerably higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, respectively, in 2% and 5% glycerol groups on both 2 and 7day after freezing(p<0.05). In 10% concentration of glycerol, the sperm seeded at each seeding temperature showed considerably higher post-thaw motility than that of non-seeding group on day 7 after freezing(p<0.01). Post-thaw viability of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}$ showed significantly higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, in 5% and 10% glycerol groups on day 7 after freezing(p< 0.05). In comparison of post-thaw motility of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed considerably higher post-thaw motility than 2% glycerol group without difference between those two groups in all seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$) on day 2 and 7 after freezing(p<0.01). In comparison of post-thaw viability of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed the same considerably higher post-thaw viability than 2% glycerol group on each thawing day(p<0.01). The canine sperm frozen and seeded by programmed freezing method showed considerably higher post-thaw motility than that frozen by direct freezing method in all different seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}$). These results indicated that the higher post-thaw motility and viability was obtained in the spermatozoa seeded than that of non-seeding, that among different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$, the sperm seeded at $-5^{\circ}C$ showed higher post-thaw motility and viability than the other temperatures, also among different concentrations fof glycerol of 2%, 5% and 10%, the sperm frozen and seeded in 5% and 10% concentration of glycerol showed higher post-thaw motility and viability than that in 2% of glycerol, and that the sperm frozen and seeded by programmed freezing method showed higher motility than that by direct freezing method.

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Application of In-Situ Mixing Hydration Accelerator on Polymer Modified Concrete for Bonded Concrete Overlay (접착식 콘크리트 덧씌우기를 위한 초속경화 첨가재 현장 혼합 폴리머 개질 콘크리트의 적용성 연구)

  • Kim, Young Kyu;Hong, Seong Jae;Lee, Seung Woo
    • International Journal of Highway Engineering
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    • v.17 no.3
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    • pp.85-95
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    • 2015
  • PURPOSES : Recently, bonded concrete overlay has been used as an alternative solution in concrete pavement rehabilitation since its material properties are similar to those of the existing concrete pavements. Deteriorated concrete pavements need rapid rehabilitation in order to prevent traffic jams on Korean expressways. Moreover, speedy and effective repair methods are required. Therefore, the use of bonded concrete overlay with ultra-rapid hardening cement has increased in an effort to reopen promptly the expressways in Korea. However, mobile mixer is required for ultra-rapid hardening cement concrete mixing in the construction site. The use of mobile mixer causes various disadvantages aforementioned such as limitation of the construction supply, open-air storage of mixing materials, increase in construction cost, and etc. In this study, therefore, hydration accelerator in-situ mixing on polymer modified concrete produced in concrete plant is attempted in order to avoid the disadvantages of existing bonded concrete overlay method using ultra-rapid hardening cement. METHODS : Bonded concrete overlay materials using ultra-rapid hardening cement should be meet all the requirements including structural characteristics, compatibility, durability for field application. Therefore, This study aimed to evaluate the application of hydration accelerator in-situ mixing on polymer modified concrete by evaluating structural characteristics, compatibility, durability and economic efficiency for bonded concrete overlay. RESULTS : Test results of structural characteristics showed that the compressive, flexural strength and bond strength were exceed 21MPa, 3.15MPa and 1.4MPa, respectively, which are the target strengths of four hours age for the purpose of prompt traffic reopening. In addition, tests of compatibility, such as drying shrinkage, coefficient of thermal expansion and modulus of elasticity, and durability (chloride ions penetration resistance, freezing-thawing resistance, scaling resistance, abrasion resistance and crack resistance), showed that the hydration accelerator in-situ mixing on polymer modified concrete were satisfied the required criteria. CONCLUSIONS : It was known that the hydration accelerator in-situ mixing on polymer modified concrete overlay method was applicable for bonded concrete overlay and was a good alternative method to substitute the existing bonded concrete overlay method since structural characteristics, compatibility, durability were satisfied the criteria and its economic efficiency was excellent compare to the existing bonded concrete overlay methods.

Probe of Unfilled Sheath of Prestressed-Concrete Girder Bridge Using Impact-Echo Method (충격탄성파법을 이용한 PC형교의 쉬이스 내부 그라우트 미충전부 탐사)

  • Lee, Sang Hun;Kim, Sang Jin;Endo, Takao;Sagara, Yuzo
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.15 no.1
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    • pp.112-119
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    • 2011
  • Sheaths are arranged in the web and lower part of prestressed-concrete girder bridges, and prestressing wires and concrete are indirectly bonded together by filling interior parts of the sheaths. However, when the filling is not sufficient, water can penetrate the interior parts of the sheaths and, thereby, prestressing wires can be corroded or transverse cracks would occur around sheaths by the freezing and thawing action of the penetrated water. Therefore, it is an important element in maintenance management of structures to find out the sheaths unfilled with grout early after construction. In this paper, in order not to damage bridge members, the impact-echo method with a new approach in application is used to probe sheaths unfilled with grout for real structures. The location of sheaths is first estimated with reinforcing bar probing instrument of radar type and the measurement locations of sheaths are determined. By sending elastic wave to the side of girders and receiving the response, the location of the unfilled part of a sheath was estimated from the difference between high frequency peak and twice the resonance frequency indicating thickness. To verify the location of void estimated by the impact-echo method, pictures were taken by an industrial fiber scope after drilling a hole.

Effect of Different Infusion Frequency of Liquid Nitrogen on Actin Filament, Mitochondria, Apoptosis and Development in Mouse 2-Cell Embryo after Freezing and Thawing (생쥐 배아 동결시 액체질소의 분사속도가 해빙후 배아의 발달, 미세섬유, 미토콘드리아 및 세포자연사에 미치는 영향)

  • 손인표;안학준;계명찬;최규완;민철기;강희규;이호준;권혁찬
    • Development and Reproduction
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    • v.4 no.2
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    • pp.161-173
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    • 2000
  • The aim of this study was to assess the effect of the frequency of the L$N_2$ infusion on the ultrastructure, metabolism and development of the embryo after freezing and thawing by computerized cell freezer. Two-cell embryos of ICR mouse were randomly allocated into fresh (control), high-frequency freezing (group 1) and low-frequency freezing (group 2). For fresh and frozen-thawed intact 2-cell embryos, total ceil number in the blastocyst was counted by fluorescent microscope after Hoechst 33258 staining. Relative amount of $H_2O$$_2$ was measured by DCHFDA. Intracellular location and membrane potential of the mitochondria were evaluated by staining with rhodamine 123 and JC-1. The structure of actin filament was also evaluated by confocal microscope. DNA fragmentation was assessed by TUNEL method after development into blastocyst. The survival rate of intact embryo was higher in group 1 than group 2 (50.7% vs. 34.6% respectively, p<0.05). The blastocyst developmental rate was significantly low in group 2 (86.7%, 76.7% vs. 44.0% for control, group 1 and group 2 respectively, p<0.05). Total cell number in the blastocyst was also significantly lower in group 2 than control (79.5$\pm$12.9, 71.6$\pm$8.0, and 62.5$\pm$4.7 for control, group 1 and group 2 respectively, p<0.05). The relative amount of $H_2O$$_2$ was higher in group 2 than other groups (15.3$\pm$3.0, 16.6$\pm$1.6 vs. 23.4$\pm$1.8, p<0.05). After JC-1 staining, relative intensity of mitochondria with high membrane potential was significantly lower in group 2 than control and group 1 (17.2$\pm$3.8, 17.4$\pm$1.3 vs. 13.2$\pm$2.0, p<0.05). In group 2, partial deletion and aggregation of the actin filament was found. DNA fragmentation rate was also hieher for group 2 versus other groups (30.8%, 36.0% vs. 65.6%, p<0.05). The frequency of the L$N_2$ infusion is an important factor for the development of frozen-thawed mouse embryo. High-frequency infusion may prevent damages of cytoskeleton and mitochondria in the embryo probably by preventing the temperature fluctuation during dehydration phase. We speculate that the application of high-frequency infusion method in human embryo may be promising.

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Systems for Production of Calves from Hanwoo(Korean Native Cattle) IVM/IVF/IVC Blastocyst I. Hanwoo IVM/IVF /IVC Blastocyst Cryopreserved by Vitrification (체외생산된 한우 배반포기배로부터 송아지 생산을 위한 체계 I. 체외생산된 한우 배반포기배의 초자화 동결보존)

  • Park, S. P.;Kim, E. Y.;Kim, D. I.;Park, N. H.;Y. S. Won;S. H. Yoon;K. S. Chung;J. H. Lim
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.349-357
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    • 1998
  • This study was to investigate whether the viability of Hanwoo IVM/IVF/IVC blastocysts was maintained after vitrification and thawing. In vitro produced Hanwoo blastocysts were vitrified by two-step method: equilibrated in EG20 for 3 min, and then exposed in EFS40 [40% ethylene glycol (EG), 18% ficoll and 10.26% sucrose in mDPBS containing 10% FBS ]and vitrified in L$N_2$for 30 - 45 sec. After thawing, in vitro survival was assessed as the re-expanded and hatched rates at 24 hand 48 h, respectively. The results obtained in these experiments were summarized as follows: From the 12 replicates, 52.5% of Hanwoo blastocysts were produced in vitro at day 7 after IVF. When the effects of freezing solution to the embryo survival were examined, there is no significant toxicity in exposure (100.0, 73.8%) compared to that af control group (100.0, 87.0%). However, when embryos were vitrified, high survival (86.2, 55.4%) was obtained although it was significantly lower than those of exposure and control group (p<0.05). When the in vitro survival of vitrified embryos according to developmental stage and culture day were examined, it showed that more advanced embryo stage exhibited a significantly higher survival rate irrespective of culture day (p<0.05). Also, even in the same development stage, the in vitro survival of day 7 embryos (re-expanded: 75.0~87.5%, hatched: 21.4~66.7%) was higher than those of day 8 embryos(re-expanded: 58.6~78.3%, hatched: 10.3~52.2%). Therefore, these results suggested that in vitro produced Hanwoo blastocysts can be successfully cryopreserved by simple two-step vitrification method using EFS40 freezing solution, particularly at the expanded and early hatching blastocyst stage regardless of embryo culture duration (day 7 or day 8 after IVF).

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Characteristics of Shear Strength and Elastic Waves in Artificially Frozen Specimens using Triaxial Compression Tests (삼축압축실험을 이용한 인공동결시료의 강도평가 및 탄성파 특성변화)

  • Kim, JongChan;Lee, Jong-Sub;Hong, Seung-Seo;Lee, Changho
    • The Journal of Engineering Geology
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    • v.24 no.1
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    • pp.111-122
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    • 2014
  • For accurate laboratory evaluations of soil deposits, it is essential that the samples are undisturbed. An artificial ground-freezing system is the one of the most effective methods for obtaining undisturbed samples from sand deposits. The objective of this study is to estimate the shear strengths and the characteristics of elastic waves of frozen-thawed and unfrozen specimens through the undrained triaxial compression test. For the experiments, Jumunjin standard sands are used to prepare frozen and unfrozen specimens with similar relative densities (60% and 80%). The water pluviation method is used to simulate the fully saturated condition under the groundwater table. When thawing the frozen specimens, the temperature is measured every minute. After the specimens are completely thawed, undrained triaxial compression tests are conducted using the same procedures as for the unfrozen specimens. During the triaxial tests (saturation, consolidation, and shear phase), compressional and shear waves are measured. The results show that the freeze-thaw process has minor effects on the peak deviatoric stress and shear strength values, and that the process does not affect the internal friction angle. The compressional wave velocity increases with increasing B-value to 1800 m/s in the saturation phase, but tends to remain constant in the process of consolidation and shearing. The shear wave velocity decreases with increasing B-value in the process of saturation, but changes velocity in accordance with the change in effective stress in the processes of consolidation and shearing. The compressional wave velocity has similar values regardless of the freeze-thaw process, but values of shear wave velocity are slighly lower in frozen-thawed specimens than in unfrozen specimens. This study is a preliminary experiment for estimating the shear strength and characteristics of elastic wave velocity in undisturbed frozen specimens that have been obtained using the artificial ground-freezing method.

Monitoring the Rate of Frozen Denaturation of Bovine Myosin by Competitive Indirect ELISA Method (Competitive Indirect ELISA를 이용한 Bovine Myosin의 동결 변성도 측정)

  • Kim, Seong-Bae;Lee, Ju-Woon;Park, Jong-Heum;Do, Hyung-Ki;Hyun, Chang-Kee;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.862-870
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    • 1998
  • This study shows the application of Ci-ELISA method for monitoring the denaturation of myosin by the frozen treatment in order to differentiate thawed beef from chilled. Hanwoo M.Semitendinosus (n=25) was treated under the two different frozen process as follows; simple frozen treatment (Exp-1) at 4 different temperatures, -10, -20, -50 and $-80^{\circ}C$, respectively, and repeated thawing-refreezing treatment (Exp-2) stored at 4 different temperatures, -10, -20, -50 and $-80^{\circ}C$, respectively. Antibodies (Abs) were produced from rabbits immunized with myosin whole molecule (MWM) isolated from beef round, heavy meromyosin S-1 (S-1) and light meromyosin (LMM) prepared by digestion of MWM. Each immunoglobulin G (IgG) was separated from antiserum. At 6 month storage, IA of anti-MWM IgG for myosin was decreased to 32.67, 32. 23, 51.52 and 34.27% in Exp-1 and to 14.82, 15.61, 25.3 and 23.7% in Exp-2 at -10, -20, -50 and $-80^{\circ}C$, respectively (P<0.05). In Exp-1, the reactivities of anti-LMM IgG were decreased to 25.12, 21.42, 49.05 and 28.96%, and those of Exp-2 were to 11.88, 9.56, 20.63 and 12.64% at -10, -20, -50 and $-80^{\circ}C$, respectively, at 6 times thawing (P<0.05). Conclusively, myosin was denaturated by freezing treatment and LMM or myosin rod part might have suffered from more extreme demage than HMM S-1, and samples at $-50^{\circ}C$ were slightly injured less than others by freezing treatment.

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