• Title/Summary/Keyword: equilibration time

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Studies on Transfer of In vitro Fertilized Mouse Embryos Following Ultrarapid Freezing III. A Study on Transfer In Vitro Fertilization Mouse Embryos Following Ultrarapid Freezing-Thawing (생쥐 체외수정난의 초급속동결 및 이식에 관한 연구 III. 생쥐 체외수정난의 초급속동결-융해란의 이식에 관하여)

  • 장규태;민관식;오석두;강대진;윤창현
    • Korean Journal of Animal Reproduction
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    • v.16 no.3
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    • pp.217-224
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    • 1992
  • These studies were carried out ot investigate on the transferred embryo development following ultrarapid frozen for 8-cell and morula of in vitro fertilization mouse embryos. The post-thaw embryo survival was evaluated and compared by cell stage of embryos and by equilibration time before ultrarapid freezing. The results obatined were summerized as follows: 1. The effects of equilibration time of 3 vs. 6 minutes before ultrarapid freezing and after thawing on the morphological survival and the viability of 8-cell and morulas embryos were not significant. 2. When the ultrarapid frozen-thawed 32 eight-cell and 33 morula embryos, and 30 fresh blastocysts were transferred to pseudopregnant recipient mice, the number of normal offsprings produced were 9(28.1%), 14(42.4%) and 18(60.0%), respectively. From the above resutls, it was concluded that the optimal conditions of pH osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOsm and 120 min., respectively,a nd somewhat high conception rate might be resulted from transfer of frozen embryos of morula stage and fresh embryos of blastocyst stage.

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Studies on Food Preservation by Controlling Water Activity 1. Measurement of Sorption Isotherm of Dried Filefish Muscle by Equilibration in Dynamic Stream of Conditioned Air (식품보장과 수분활성에 관한 연구 1. 조절기류에 의한 건조말쥐치육의 등온흡습곡선의 측정)

  • HAN Bong-Ho;CHOI Soo-Il
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.14 no.4
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    • pp.189-193
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    • 1981
  • An apparatus for continuous measurements of sorption isotherm of dried food was manufactured to shorten the time required for equilibration. The apparatus was so designed that the temperature, air velocity and relative humidity in the experimental chamber could be controlled. The use of dynamic stream of conditioned air with a velocity of 0.2m/sec, instead of static atmosphere, allowed a faster equilibration of dried filefish muscle at $25^{\circ}C$. The mean time necessary for the equilibration of dried filefish muscle at the water activity of a given state to a higher water activity was about 45 hours. The monolayer moisture content of dried filefish muscle calculated from BET-equation was 0.092 kg water /kg dry matter at $25^{\circ}C$.

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Studies on the Factors Influencing Survival Rates of Frozen Bovine Demi-Embryos (소 동결분할배의 생존선에 영향을 미치는 요인에 관한 연구)

  • 김상근;남윤이;이만휘;현병화
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.287-292
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    • 1997
  • This study was carried out to investigate the effects of concentration and kinds of cryoprotectants, equilibraction time, thawing temperature and time, sucrose concentration on the survival rates of frozen bovine demi-embryos. The bovine demi-embryos following dehydration by cryoprotectants a various concentration of sucrose were freezed by cell freezer and thawed in 3$0^{\circ}C$ water bath. Survival and in vitro developmental rates was defined as development rates on in vitro culture or FDA-test. The results are summarized as follows : 1. The high survival rates of demi-embryos after frozen-thawing in freezing medium was attained 2.0M glycerol. The high survival rates of demi-embryos after frozen-thawing in freezing medium was obtained using single cryoprotectant(25.0~30.0%) than mixed cryoprotectants(16.7~19.0%). 2. The survival rates of demi-embryos after frozen-thawing in freezing medium added 1.5M, 2.0M glycerol+0.25M sucrose(37.5~33.3%) were higher survival rates than those of sucrose concentration of 0.50, 0.75M(12.5~26.7%). 3. The equilibration time on the survival rates of demi-embryos was attained after short period of time(30.0~35.0%) in the freezing medium higher than long period of time(21.1%). 4. The thawing temperature on the survival rates of demi-embryos was attained at 3$0^{\circ}C$ of thawing temperature(26.7~40.0%) higher than $25^{\circ}C$ or 37$^{\circ}C$ of thawing temperature(13.3~20.0%). 5. The thawing time on the survival rates of demi-embryos was attained at 1~5 minutes of thawing time(26.7~33.3%) in the freezing medium higher than 10 minutes of thawing time(13.3~18.8%).

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Studies on the Survival and In Vitro Fertilization Rates of Frozne-Thawed Porcine Embryos and Immature Oocytes (돼지 수정란 및 미성숙 난자의 동결융해 후의 생존율에 관한 연구)

  • 김상근;이명헌;남윤이
    • Korean Journal of Animal Reproduction
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    • v.22 no.2
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    • pp.187-194
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    • 1998
  • In order to determine suitable conditions for rapid freezing of porcine embryos, the kind and concentration of cryoprotectants, sucrose concentrations, equilibration time and thawing temperature in freezing medium were examined in relation to the survival of frozen-thawed oocyte and embryos. The results obtained are as follows : 1. The suitable concentrations of cryoprotoctant in the freezing medium which consisted of TCM-199+20% FCS were 1.5M for glycerol, 2.0M for DMSO, 2.5M for ethylene glycol, and 2.0M for propanediol. The sucrose concentration of 0.25M in the medium was found to optimal because the survival rate was markedly higher at this concentration when compared to the others. The survival rate was relatively high when the frozen embryos were thawed at 30$^{\circ}C$ in the freezing medium containing 2.5M cryoprotectants. The equilibration periods of 2.0 and 5.0 minutes revealed the higher survival in the media containing 1.5 or 2.1M glycerol when compared to 10 and 15 minutes. 2. The fertilization rates of frozen-thawed follicular oocytes which matured in vitro for 1, 12, 24 and 48 hours were 6.7~26.7% depending on the maturation time, and the rates were relatively high for those matured for a short period of time. The survival rates of frozen-thawed oocytes which matured in vitro for certain periods and fertilized were 10.0~30.0% depending on the maturation time.

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Sperm Cryopreservation of Tetraploid Pacific Oysters Crassostrea gigas (4배체 참굴(Crassostrea gigas)의 정자 동결보존)

  • Park, Mi Seon;Min, Byung Hwa;Lim, Hyun-Jeong;Hur, Young Baek;Do, Yong Hyun;Myeong, Jeong-In
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.6
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    • pp.853-859
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    • 2014
  • The goal of this study was to evaluate the effects of cryoprotectants [dimethyl sulfoxide (DMSO), methanol, polyethylene glycol, and propylene glycol], cryoprotectant concentrations (10% and 20%), equilibration time (3, 10, and 30 min), cooling rate ($3^{\circ}$, $5^{\circ}$, $7^{\circ}$, and $10^{\circ}/min$), and straw size (0.25 and 0.5 mL) for sperm cryopreservation of tetraploid Pacific oysters. There was a significant difference among the four cryoprotectants, with 10% DMSO yielding the highest post-thaw survival and activity index of sperm. A significant relationship was observed between the cryoprotectant and its concentration. The sperm with equilibration times of 30 min yielded higher post-thaw survival and activity indices than those with 3 and 10 min equilibration times. The sperm cooled at a rate of $5^{\circ}/min$ yielded the highest post-thaw survival and activity index, and the results were significantly different from those observed for cooling at $7^{\circ}$ and $10^{\circ}/min$. Post-thaw survival and activity indices of sperm using a 0.25-mL straw were significantly higher than those using a 0.5-mL straw.

Studies on the Survival Rates after Ultrarapidly Frozen-Thawing of Porcine Embryos (돼지 수정난의 초급속 동결 융해후의 생존성에 관한 연구)

  • 김상근;이봉구
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.125-131
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    • 1992
  • This study was carried out to investigate the effects of concentration and equilibration time of cryoprotective agents on the survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rat to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5, or 4.0M glycerol was 65.3, 61.8, 64.3, 59.4 or 39.4%, respectively. Addition of 0.25M sucrose into the freezing medium containing 2.0M glycerol showed higher survival rate than those of 2.5~4.0M glycerol. 2. The survival rates of porcine embryos after ultraradpid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0, 2.5, 3.0, 3.5 or 4.0M DMSO was 65.6, 67.6, 68.6, 60.6 or 23.6%, respectively. However, addition of 0.25M sucrose into the freezing medium containing 3.0M DMSO showed higher survival rate than those of 2.0, 2.5, 3.5 or 4.0M DMSO. 3. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5 or 4.0M propanediol was 63.2, 60.3, 62.1, 52.3 or 24.3%, respectively. Addition of 0.25M sucroese into the freezing medium containing 2.0M propanediol showed higher survival rate than those of 2.5~4.0M glycerol. 4. The survival rates of porcine embryos after ultrarapid frozen-thawing the freezing medium of 2.0M glycerol added 0.10, 0.25, 0.50 or 0.75M sucrose was 61.8, 70.8, 67.6 or 52.2%, respectively. Addition of 2.0M glycerol into the freezing medium containing 0.25M sucreose showed higher survival rate than that those of 0.10, 0.50 or 0.75M sucrose. 5. The higher suvival rate of porcine embryos were attained at short period of equilibration time 92.5~5min.) in the freezing medium added 0.25M sucreose and 3.0M compared to those of 10 or 20min. equilibration time in the same condition.

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Studies on the Survival and In Vitro Fertilization Ratesof Frozen-Thawed Bovine Immature Oocytes (소 동결 미성숙 난자의 생존율과 체외수정율에 관한 연구)

  • 김상근;유상식;김용섭
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.207-211
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    • 1998
  • 본 연구는 소 미성숙 난자의 동결에 있어서 미성숙 난자의 1, 2, 4, 8 시간 성숙배양한 다음 동결 융해 후 체외배양하였을 때 체외수정율과 생존율을 조사하고저 수행하였다. 미성숙 난자들 1∼8시간 성숙배양후 동결융해 및 체외수정시켰을 때 수정율은 12.8∼30.9%로서 단시간의 체외성숙 배양이 높게 나타났으며(23.8∼28.8%), 또한 체외배양시 생존율은 11.8∼29.9%를 나타냈다.

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Sperm Cryopreservation of Pacific Cod Gadus macrocephalus (대구(Gadus macrocephalus)의 정자 동결보존)

  • Do, Yong Hyun;Cho, Jae Kwon;Lee, Hee Jung;Min, Byung Hwa
    • Korean Journal of Ichthyology
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    • v.31 no.4
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    • pp.195-200
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    • 2019
  • An experiment was performed to obtain cryopreservation techniques of Pacific cod Gadus macrocephalus sperm. Milt were cryopreservation using five cryoprotectant demethyl sulfoxide (DMSO), ethylene glycol (EG), glycerol, methanol and propylene glycol (PG) with marine fish ringer's solution (MFRS) as diluent. Milt were cryopreserved each experimental methods like cryoprotectants (10% and 20%), equilibration time (3, 5 and 10 min) and freezing protocols (liquid nitrogen vapor above 3, 8 and 12 cm). Post-thaw sperm survival rate revealed the highest in 10% PG with optimum methods of equilibration time (3 min) and freezing protocol (liquid nitrogen vapor above 8 cm) about 21.3±1.8%. Hatching rate of fertilization eggs using fresh and cryopreserved sperm were no significantly different.

Fertilization In vitro of follicular oocytes and cryopreservation of embryo fertilized and developed In vitro In Korean native cattle (한우 난포란의 체외수정 및 체외수정란의 동결보존에 관한 연구)

  • Choe, Sang-yong;Kong, Ill-keun;Joo, Young-kuk;Rho, Gyu-jin;Kim, Yong-kweon;Park, Choong-saeng
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.757-762
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    • 1993
  • The ovaries of Korean Native cows or heifers were obtained from an abattoir and kept on 20 to $25^{\circ}C$ and transported to laboratory within 2 hrs. The follicular oocystes were collected from 2~6mm follicles in diameter and classified into 3 grades by the morphology of cumulus cells attached. The oocytes were matured in vitro(IVM) for 24 hrs. in TCM-199 supplemented with $23{\mu}g/ml$ FSH, $10{\mu}g/ml$ LH, $1{\mu}g/ml$ estradio-17 ${\beta}$ and granulosa cells at $39^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro(IVF) by incubation for 12 hrs. of epididymal spermatozoa pretreated with heparin, and then the zygotes were co-cultured in vitro(IVC) with oviductal epithelial cells for 7 to 9 days. Assessment of maturation revealed that 93.0%(147/158) of grade I oocytes had expanded of cumulus cells, which was higher(p<0.05) than the 79.4%(85/107) of grade II oocytes. Compared to epididymal sperm(32.9%), the insemination with frozen and thawed sperm resulted in slightly lower(20.5%), but not significant, development to morulae and blastocysts from grade I oocytes. Co-culture of bovine IVF embryos with oviductal epithelial cells improved the development to transferable embryos significantly(38.1%), compared to co-culture with granulosa cells(20.0%). When VF bovine embryos were vitrified at blastocyst, the post-thaw survival rate was obtained higher resulf for 1 min. equilibration time(82.6%) or 2 min.(73.9%) than 3 min.(18.2%) in EFS solution.

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Moringa Oleifera, A Biosorbent for Resorcinol Adsorption-Isotherm and Kinetic Studies

  • Kalavathy, M. Helen;Swaroop, G.;Padmini, E.;Lima Rose, Miranda
    • Carbon letters
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    • v.10 no.1
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    • pp.23-32
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    • 2009
  • The adsorption of phenolic compound resorcinol on activated carbons prepared from Moringa oleifera (Drumstick bark) has been investigated. Activated carbon was prepared by impregnating Moringa oleifera with 50% phosphoric acid in the ratio of 1:1 and 1:2(w/w), designated as MOAC1 and MOAC2. Equilibrium and isotherm studies were carried out. The influences of variables such as contact time, initial concentration of resorcinol, carbon dosage in the solution on percentage adsorption and adsorption capacity of the bark have been analysed. The equilibration time was found to be 4 h. Kinetics of resorcinol onto activated carbons was checked for pseudo first order and pseudo second order model. It was found that the adsorption of resorcinol follows pseudo second order kinetics for both MOAC1 and MOAC2. The isotherm data were correlated with isotherm models, namely Langmuir and Freundlich. Adsorption isotherms were satisfactorily fitted by both the Langmuir and Freundlich model for MOAC1 and MOAC2.