2단계 급속동결 및 초자화 동결한 토끼상실배의 체외생존성에 관한 연구

정구민,이창규,임경순,김수헌 서울대학교농과대학농업개발연구소 1990 서울대농학연구지 Vol.15 No.1

본 시험은 수정란 급속동결보존기술의 개발을 위한 기초자료를 얻기 위하여 2단계 급속동결 및 초자화동결이 토끼 상실배의 체외발생등에 미치는 영향을 검토하고자 실시하였으며, 얻어진 결과는 다음과 같다. 1. 토끼 상실배를 1.5, 2.0, 2.5 및 3.0 M glycerol과 0.5 M sucrose가 포함된 동결액에 실온에서 10분간 노출후 -30℃에서 30∼40분간 정치하여 급속동결하였을 때 발생율은 각각 36.4, 83.3, 92.3 및 84.2%로 2.5 M glycerol에서 가장 높았다. 2. 토끼 상실배를 1.5, 2.0, 2.5 및 3.0 M 1,2-propanediol과 0.5 M sucrose가 포함된 동결액에 실온에서 10분간 노출 후 -30℃에서 30∼40분간 정치하여 급속동결하였을 때 발생율은 각각 26.6, 55.6, 65.0 및 52.9%로 2.5 M 1,2-propanediol에서 가장 높았으나, glycerol을 사용했을 때 보다 낮았다. 3. 토끼 상실배를 2.5, 3.0 및 3.5 M glycerol과 0.5 M trehalose가 포함된 동결액에 실온에서 10분간 노출후 액체질소에 침지하여 초급속동결한 결과 회수율은 각각 87.5, 92.5 및 92.5%, 형태적으로 정상인 수정란의 비율은 각각 37.5, 55.5 및 60.0%, 그리고 발생율은 각각 13.3, 36.4 및 37.5%로 3.5 M glycerol에서 가장 높았으나 초자화동결법보다 현저하게 낮았다. 4. 토끼 상실배를 25% glycerol과 25% 1,2-propanediol을 함유한 동결액에 실온에서 10분간 노출후 초자화동결했을 때 발생율은 75.0%로 실온에서 형평한 후 초자화동결이 가능하였다. This experiment was carried out to investigate on in vitro development of rabbit monla frozen by 2-step feezing and vitrification. The results obtained from this experiment are as follows; 1. When rabbit morula in m-PBS containing 1.5, 2.0, 2.5 or 3.0 M glyceral and 0.5 M sucrose for 10 min at room temperature were cooled at -30℃ for 30 to 40 min and plunged into liquid nitrogen, the proportion of embryo developed to expanded blastoyst was 36.4, 83.3, 92.3 and 84.2%, respectively. Glycerol 2.5 M showed higher survival than others. 2. When rabbit morula in m-PBS containing 1.5, 2.0, 2.5 or 3.0 M 1, 2-propanediol and 0.5 M sucrose for 10 min at room temperature were cooled at -30℃ for 30 to 40 min and plunged into liquid nitrogen, the proportion of embrye developed to expanded blastocyst was 26.6, 55.6, 65.0 and 52.9%, respectively. 1, 2-propanediol was less effective than glycerol. 3. When rabbit morula in m-PBS containing 2.5, 3.0 or 3.5 M glycerol and 0.5 M trehalose for 10 min at room temperature were plunged into liquid nitrogen and thawed rapidly, the recovery rate of embryo was 87.5, 92.5 and 92.5%, the proportion of morphologically normal embryo was 37.5, 55.5 and 60.0%, and the proportion of embryo developed to expanded blastocyst was 13.3. 36.4 and 37.5%, respectively. The proportion of embryo developed to expanded blastocyst was higher in vitrification than in plunging into liquid nitrogen. 4. When rabbit morula were frozen by vitrification in m-PBS containing 25% glycerol. 25% 1, 2-propanediol, the proportion of embryo developed to expanded blastocyst was 75.0%, the result suggested that rabbit embryos could be frozen by vitrification after equilibration at room temperature.

Improved production of phleichrome from the phytopathogenic fungus Cladosporium phlei using synthetic inducers and photodynamic ROS production by phleichrome

So, K.K.,Jo, I.S.,Chae, M.S.,Kim, J.M.,Chung, H.J.,Yang, M.S.,Kim, B.T.,Kim, J.K.,Choi, J.K.,Kim, D.H. Society for Bioscience and Bioengineering, Japan ; 2015 Journal of bioscience and bioengineering Vol.119 No.3

Two different diketopiperazines, cyclo-(l-Pro-l-Leu) and cyclo-(l-Pro-l-Phe), which were isolated from the culture filtrate of Epichloe typhina and found to be inducers of phleichrome production, were chemically synthesized and evaluated for use in the improved production of phleichrome from wild-type and UV-mutagenized strains (M0035) of Cladosporium phlei. When supplemented with PDA and V8 juice agar media, both inducers showed significant increases in the production of phleichrome. Phleichrome production was increased in a dose-dependent manner up to a concentration of maximum yield for both inducers. No further significant induction was observed by supplementing inducers over the concentration of maximum yield. Among the two inducers, cyclo-(l-Pro-l-Phe) showed better inducing capability than cyclo-(l-Pro-l-Leu). The maximum yield was observed from the M0035 strain grown on V8 juice media supplemented with 150 μM cyclo-(l-Pro-l-Phe), which was estimated to be 232.6 mg of phleichrome per gram of mycelia and 10.2 mg of secreted phleichrome per 20 agar-plugs. Interestingly, growth inhibition was observed on V8 juice agar media with 100, 150, and 200 μM cyclo-(l-Pro-l-Phe) but not on PDA with the same amount of inducer, which suggests that the inhibitory effect might be through the overproduction of phleichrome rather than the toxic effect of the inducer itself. Superoxide production by purified phleichrome was dramatically stimulated upon illumination, thus demonstrating photodynamic production of superoxide in vitro by phleichrome.

유우개량을 위한 유단백질의 유전적 다형에 관한 연구 Ⅰ. αS1 - Casein 및 K - Casein 의 유전적 변이체

한상기(Sang K . Han),이기만(Khy M . Lee),정의용(Eui Y . Chung),장경진(Kyung J . Jang) 한국축산학회 1984 한국축산학회지 Vol.26 No.3

Genetic variants of αs₁-casein (αs₁-Cn) and K-casein (k-Cn) in milk proteins from 138 individual Holstein cattle in Korea were investigated by means of strach-gel-urea electrophoresis and the appearance of phenotypes, gene frequencies and genetic similarities were compared with those of foreign Holstein and of her breeds. The results are summarized as follows: 1. The milk proteins in cattle were controlled by codominant autosomal allelic genes of 9αs₁-Cn^A and αs₁-Cn^B(2 types), and K-Cn^A and K-Cn^B (2 types). 2. The distribution of phenotypes in each milk protein loci was αs₁-Cn BB 124, BC 14; K-Cn AA 76 Ab 49, BB 13. The number of phenotypes coxesponded closely to expectation of Hardy-Weinberg equilibrium (αs₁-Cn:0.7 $gt;P$gt;0.5, k-Cn;0.7$gt;P$gt;0.5). 3. Gene frequencies for genetic variants of the milk protein loci were analyzed. By the predominant allele in αs₁-Cn was αs₁-Cn B with a frequency of 0.949, whereas αs₁-CnC was in low gene frequency (0.051). Among the two alleles for K-Cn, K-Cn A gene showed to occur in high frequency (0.728). However, the Kin B was in low gene frequency (0.272).

生體內 藥劑耐性 및 Plasmid 傳達에 關한 硏究

金鎬薰,李明遠,李英姬,金基湘,兪天權,吳明燉,李悌煥,李点揆,琴東吉,鄭泰華,李潤台 국립보건연구원 1989 국립보건원보 Vol.26 No.

사과 矮性臺木 繁殖 및 育苗에 關한 硏究

金聲遠,李庚熙,鄭大敎,金炳友,鄭文在 건국대학교 1973 學術誌 Vol.15 No.1

For the supply of the dwarf root stock of apple tree rapidly, there must be proper method in propagation of it, therefore we purposed to find out the profitable method of propagation of dwarf apple root stock and the influence of root stock on the plant division and the growth of apple tree scions. In this study, 4 kinds of common root stock, 5 varieties of dwarf apple tree, and 6 kinds of spur variety were employed, the result were as follows. 1. Layer treatment was suitable on the division and growth of MM 106, MM 111, M 7, and especially MM 111 was promoted on root formation. 2. At mound treatment, M 7 produced most divided plant than the others, and promotion of root formation was expected on MM lines. 3. Comparing of plant division between layer treatment and mound treatment, layer treatment was more effective than mound treatment, but root formation was not. 4. For the propagation of dwarf apple tree by the grafting, M. baccata and M. sieboldii was profitable as the root stock of dwarf apple tree, because the union by callusing and the growth after the grafting was effective by using of those root stock. 5. Considering the growth of scion of apple tree, MM 106 was more suitable than M 9 as the inter media stam, and the grafting of Mac spur on the inter media stem, MM 106, most was promoted in growing and rooting. 6. There were different combination on root stock and scion for the propagation of spur apple tree, proper combination were as follows, Sundale Sturdee spur G.D. …… M. prunifolia, Nugget spur Y.D.ㆍSky spurㆍMac spur,ㆍJonathan…… M. sieboldii, Wayne spurㆍNugget spur,…… M. baccataㆍM. sieboldii.

Glucose와 Phosphate가 제거된 M-TALP 배지에서의 난구세포 공배양에 의한 임신율 향상에 관한 연구

정범식,장우현,이문희,김지연,방지호,김규현,서태광,Chung, B.S.,Chang, W.H.,Lee, M.H.,Kim, J.Y.,Bang, J.H.,Kim, K.H.,Suh, T.K. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.1

The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

In Vitro/In Vivo Development after Thawing of Vitrified Mouse Blastocysts by Culture Condition and Embryo Transfer Method

김묘경,김은영,이봉경,윤산현,박세필,정길생,임진호,Kim, M.K.,Kim, E.Y.,Yi, B.K.,Yoon, S.H.,Park, S.P.,Chung, K.S.,Lim, J.H. The Korean Society for Reproductive Medicine 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

본 실험은 초자화 동결된 생쥐 배반포기배의 융해 후 배양조건 및 이식방법이 난자의 생존에 미치는 효과를 조사하고자 실시하였다. 체외수정후, M16배양액에서 4일동안 배양하여 얻어진 생쥐 배반포기배는 EFS40 (40% ethylene glycol, 18% Ficoll, 0.5 M sucrose가 함유된 PBS)으로 초자화동결하였다. 실험 I에서는 융해 후 배양조건에 따른 난자들의 체외/체내 생존율을 조사하였다. 융해된 난자가 M16과 4 mg/ml 소혈청알부민과 20 가지 아미노산이 함유된 m-CR1 (2% BME 아미노산 용액, 1% MEM 아미노산 용액) 및 단층배양이 유도된 난구세포 (10% FBS가 함유된 m-CR1배양액)에서 각각 배양되었을 때, 융해 후 24시간째 체외 생존율은 배양조건에 따라 차이가 없었다(75.6, 83.1, 82.4%). 그러나 체내 발달율에 있어서 임신 15일째 생존 산자율은 39.0, 49.0, 38.1%로서 유사한 성적을 나타냈으나, 전체 착상율에 있어서는 m-CR1 (80.4%)에 배양되었을 때, M16 (51.2%), 난구세포와 공배양시 (57.1%)보다 유의하게 높은 생존율을 보였다(p<0.05). 실험 II에서는 수정란 이식 방법에 따른 체내 발달율을 조사하였다. 배반포기배를 융해 후 체외배양없이 곧바로 가임신 2, 3일째 대리모에 이식을 실시하였을 때, 가임신 2일째 대리모에서는 임신징후를 얻지 못하였고, 가임신 3일째 대리모에서는 50.0%의 착상율과 15.4%의 정상산자율을 얻었다. 그러나, 이러한 결과는 융해 후, 16시간 배양하여 가임신 3일째 대리모에 이식 (73.5, 57.1%)하는 경우보다 유의하게 낮은 결과였다(p<0.05). 실험 III에서는 초자화 동결된 배반포기배의 융해 후 배양시 발달이 늦어진 수정란의 이용효율을 극대화시키기 위해 융해한 4일째 초기, 5일째 초기, 5일째 팽창 배반포기배의 체외/체내 생존율을 조사하였다. 가장 높은 체외 생존율은 5일째 팽창 배반포기배 (78.3%)에서 얻었으나, 체내 발달율 (산자율, 착상율)에 있어서는 4일째 초기 배반포기배 (33.3, 66.7%)의 경우가, 5일째 팽창 배반포기배(29.0, 38.7%)의 경우보다 높았다(p<0.05). 따라서 본 연구의 결과는 배양조건과 수정란 이식방법에 따라 초자화 동결된 배아의 체외/체내 발달율을 높일 수 있으며, 발달이 늦은 배반포기배의 체내 발달율은 체외 배양시간이 길어질수록 낮아짐으로, 5일째 팽창 배반포기배보다 4일째 초기 배반포기배를 동결하는 것이 더 유용하다는 것을 알 수 있었다. This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.

抗卵丘細胞 抗體가 牛卵胞卵의 體外成熟과 受精 및 豚發達에 미치는 影響

鄭吉生,朴世必,鄭炯敏,嚴基鵬,李勳澤,鄭炳鉉 건국대학교 동물자원연구센터 1991 動物資源硏究誌 Vol.16 No.-

本 硏究에서는 卵巢內에 존재하는 다수의 未成熟 卵胞卵을 회수한 다음 이를 in vitro system에서 成熟, 受精 및 胚發達에 있어서 卵丘細胞(cumulus cell)가 미치는 영향과 그 기전을 조사하였다. 卵丘細胞에 대하여 좀더 구체적인 검토를 수행할 목적으로 牛卵胞卵에 대한 抗 卵丘細胞 抗體(Rabbit anti-bovine cumulus cell antibody)를 제조하여 Ammonium sulfate precipitation과 Protein A-sepharose CL-4B affinity chromatography의 의해 IgG만을 분리 정제한 다음, indirect ELISA와 Immunoprecipitation法에 의해 확인된 이들 각각의 抗體(intact cumulus cell antibody, solubilized cumulus cell antibody)를 牛卵胞卵의 體外成熟과 受精時 첨가하는 卵丘細胞가 體外成熟과 受精 및 갑發達에 미치는 영향을 조사하여 다음과 같은 결과를 얻었다. 1) 無處理 卵丘細胞와 溶解 卵丘細胞를 抗原으로 처리한 개체로부터 血淸을 채취하여 間接酸素免疫分析法으로 抗體 형성여부를 확인한 결과 1:12,800의 稀釋倍率에서도 두 抗血淸 處理區가 對照 血淸區에 비해 높은 O.D.價를 나타냄으로써 두 가지 형태의 抗原이 다같이 抗卵丘細胞 抗體를 생산하는 것으로 확인되었다. 2) Iodogn과 Immunoprecipitation를 이용하여 항체의 생성을 재차 確認하였는데 Iodogen method를 이용한 牛卵丘細胞의 ?? Iabelling은 25,000/10㎕ cmp이었다. 또한 ??-bovine cumulus cell과 각각의 抗血淸 反應 결과 생성된 Immunoprecipitates의 radioactivity와 O.D價를 조사하였던바, 無處理 혹은 溶解化된 卵丘細胞에서 공히 높은 抗體의 생성을 확인했으며 두 處理群을 비교해 볼 때 溶解化된 卵丘細胞 抗體가 無處理 卵丘細胞 抗體보다 더 높은 反應性과 O.D價를 가지고 있음을 알 수 있었다. 3) 無處理 卵丘細胞에 대한 抗血淸과 溶解化된 卵丘細胞에 대한 抗血淸으로부터 抗體를 분리해 냈을 때 정제된 Ig G의 O.D.價를 각각 1.19mg/ml 이상이었다. 4) 卵丘細胞가 부착된 卵胞卵과 제거된 卵胞卵에 無處理 卵丘細胞에 의해 생성된 抗體를 처리한 결과, Metaphase Ⅱ 段階에까지 발달한 卵胞卵은 卵丘細胞가 제거된 卵胞卵의 對照區를 포함한 모든 抗體 處理區의 경우 47.6∼59.1%로써 卵丘細胞가 부착된 卵胞卵의 對照區 78.8%에 비해 유의하게 낮았다. (P<0.01). 5) 卵丘細胞가 부착된 卵胞卵과 제거된 卵胞卵의 溶解 卵丘細胞에 의해 생성된 抗體를 처리한 결과, M Ⅱ 단계에까지 발달한 卵胞卵은 卵丘細胞가 제거된 卵胞卵의 對照區를 포함한 모든 抗體 處理區의 경우 47.6∼59.1%로써 卵丘細胞가 附着된 卵胞卵의 對照區 82.1%에 비해 有意하게 낮았다.(P<0.01). 抗卵丘細胞 抗體의 작용으로 인하여 이러한 卵丘細胞의 효과는 억제되는 결과로 미루어 볼 때 卵丘細胞는 體外成熟시 卵胞卵의 核成熟을 증진시키는 것으로 사료된다. 6) 卵丘細胞가 부착되었거나 또는 제거된 成熟 牛卵胞卵에 武處理 卵丘細胞에 대한 抗體를 처리한 다음 體外受精을 실시한 결과 卵丘細胞가 제거된 卵胞卵에 있어서 對照區를 포함한 모든 抗體 處理區의 受精率 45.0∼53.7%는 卵丘細胞가 부착된 卵胞卵에 있어서 對照區의 受精率 64.3%에 비해 다소 낮았다. 한편 雌性 및 雄性 前核이 동시에 형성된 牛卵胞卵의 비율은 卵丘細胞를 제거한 卵胞卵의 對照毆를 포함한 모든 抗體 處理區가 18.2∼38.9%로써 卵丘細胞가 부착된 卵胞卵에 있어서의 對照區 51.9%에 비해 有意하게 낮았다.(P<0.05). 7) 卵丘細胞가 부착되었거나 또는 제거된 成熟 卵胞卵에 溶解 卵丘細胞에 대한 抗體를 처리한 다음 體外受精을 실시한 결과 卵丘細胞가 제거된 卵胞卵에 있어서 對照區를 포함한 모든 抗體 處理區의 受精率 45.0∼52.5%는 卵丘細胞가 부착된 卵胞卵에 있어서 對照區의 受精率 62.8%에 비해 다소 낮았다. 또한 雌性 및 雄性 前核이 동시에 형성된 卵子의 비율은 卵丘細胞가 제거된 卵胞卵에 있어서의 對照區를 포함한 모든 부착된 卵胞卵가 26.3∼38.3%로써 卵丘細胞가 부착된 卵胞卵에 있어서의 對照區 55.6%에 비해 有意하게 낮았다.(P<0.05). 8) 抗體處理를 받은 牛卵胞卵의 체외발생을 조사하였던 바, 卵丘細胞가 제거된 卵胞卵에 있어서 對照區를 포함한 모든 抗體 處理區의 受精率이 8-細胞期, 16-細胞期 및 桑實胚期 혹은 胚盤胞期까지 발달한 비율은 7.1∼14.5, 2.9∼5.9 및 1.5%∼2.9%로써 卵丘細胞가 부착된 난자에 있어서 對照區의 18.6, 10.0 및 8.6%에 비해 낮은 胚發生率을 나타냈다. 抗卵丘細胞 抗體를 처리함으로써 卵丘細胞 작용이 억제된다는 本 硏究 결과는 卵丘細胞가 卵胞卵의 體外受精率과 前核形成率 및 胚發生能에 유의한 효과가 있다는 것을 시사한다. These experiments were carried out to investigate the effect of cumulus cells on in vitro maturation, fertilization, and subsequent development of follicular oocytes recovered from the bovine ovaries as well as to study the mechanisms involved in these processes. To examine the specific role of cumulus cell, antisera to bovine cumulus cell were produced Japanese Giant rabbit by repeated immunization of intact or solubilized bovine cumulus cell and purified IgG by the Ammonium sulfate precipitation and Protein A-Sepharose affinity chromatography. Then, these IgG of the bovine cumulus cell-specific antibodies were confirmed by indirect ELISA and Immunoprecipitation, and the effects of theses antibodies were investigated in vitro maturation, fertilizaiton, and development of bovine follicular oocytes. The results obtained in these experiment were summarized as follows: 1. The titer of the antibodies to cumulus cell determined by indirect ELISA using intact or solubilized bovine cumulus cell coasted plated was very high in both intact and solubilized cumulus cell. Especially, the optical density(O.D.) at 1: 12,800 dilution of antibodies was significantly higher than those of non-immunized control sera. 2. To reexamine the immunity of antibody, cumulus cells were labelled with I using the Iodogen method and the radioactivity of I into cumulus cell was shown approximately 25,000cpm/10㎕. The high radioactivity and O.D. value in immunoprecipitating reaction of I-bovine cumulus cell with anti-cumulus cell antisera were reconfirmed the existence of immuno-reactivity of antibody to cumulus cell. In addition, reactivities and O.D. values of solubilized cumulus cell antibodies were significantly higher than those of intact cumulus cell antibody. 3. O.D. values of the purified intact and solubilized cumulus cell antibodies were more than 1.19mg/ml. 4. When the follicular oocytes were treated with antibody to intact cumulus cell, the maturation rates of cumulus-enclosed and removed oocytes were ranged from 47.6 to 59.1%. These values are significantly lower than those(78.8%) fo follicular oocytes cultured without the antibody (p<0.01). 5. The maturation rates of cumulus-enclosed and removed oocytes treated with antibody against solubilized cumulus cell were ranged from 46.7 to 59.1%, showing significantly lower than those(82.1%) of oocytes cultured in antibody free medium(P<0.01). These results that the beneficial effect of cumulus cell to the oocyte maturation was inhibited by the action of antibody to cumulus cell suggest that cumulus cell enhances nuclear maturation of follicular oocytes. 6. When the follicular oocytes matured in vitro were treated with antibody to intact cumulus cell, the fertilization rates of cumulus intact and removed oocytes were ranged from 45.0 to 53.7%. These values are slightly lower than those(64.3%) of follicular oocytes not treated with the antibody, and increased frequencies of both male and female pronucler formations were detected in cumulus intact oocytes cultured in medium without antibody (P<0.05). 7. The fertilizaiton rates of cumulus intact and removed oocytes treated with antibody to solubilized cumulus cell were ranged from 45.0 to 52.5%, significantly lower than those(62.8%) of oocytes cultured in antibody free medium, and increased frequencies of ova with male and female pronuclei formations were found in the presence of cumulus cells(P<0.05). 8. The development rates of cumulus intact and removed oocytes to 8-, 16-cell, and morula or blastocyst stage after treatment of intact and solubilized cumulus cell antibody were ranged from 7.1 to 14.5 from 2.9, and from 1.5 to 2.9T, respectively. These rates were slightly lower than 18.6, 10.0 and 8.6% of cumulus intact oocytes cultured in medium without the antibody. The results that beneficial effects of cumulus cell to the pronuclear formation and embryo development were blocked by the action of antibody to cumulus cell suggest that cumulus cell significantly promotes normal fertilizaiton with proper pronuclear formation and embryo development.

Machining of the KSTAR TF coil structure

K. H. Hong,C. H. Choi,D. H. Park,D. K. Hur,H. Sin,H. K. Park,H. T. Kim,J. H. Won,J. H. Lim,J. M. Chung,박주식,J. W. Sa,J. Y. Song,N. G. Kim,S. H. Hwang,Y. D. Chu,Y. G. Song 한국물리학회 2006 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.49 No.III

The KSTAR Toroidal Field (TF) magnet system consists of 16 TF structures. Each structure must be machined for precise assembly and reducing error field. Dimensions of the structure are a height of 4.5 m and a width of 3.2 m. The allowable tolerance of the structure is ±1 2 mm, and that of the bolt hole for Poloidal Field (PF) coil basement is ±0.5 mm. The final machining of the structure is also important for assembly of the interface structures such as toroidal ring, PF coil structures, and Central Solenoid (CS) structure. Before coil encasing, the inside surface of the structure is machined. Most of the outer surface of the structure, except for the surface of the intercoil structure (ICS), is machined after final welding. In particular, the side surface of an inboard leg, the connection plate of the ICS, the toroidal ring basement, the PF structure basements, shear key holes, and conical bolt holes are precisely machined. Fabrication and assembly tolerances are absorbed by the thickness of the electrical insulation, the spacer of the shear key, and the taper ring of the conical bolt. Six TF coil structures have been assembled and two structures are waiting for assembly. Another three structures are under final machining. A laser tracker is used for three dimensional survey and measurement of the structure.

Continuous lithium mining from aqueous resources by an adsorbent filter with a 3D polymeric nanofiber network infused with ion sieves

Chung, W.J.,Torrejos, R.E.C.,Park, M.J.,Vivas, E.L.,Limjuco, L.A.,Lawagon, C.P.,Parohinog, K.J.,Lee, S.P.,Shon, H.K.,Kim, H.,Nisola, G.M. Elsevier 2017 Chemical Engineering Journal Vol.309 No.-

Electrospun composite nanofiber (NF) was fabricated and employed as an adsorbent membrane filter in a continuous Li<SUP>+</SUP> mining process from seawater. The filter was composed of a hydrophilic polyacrylonitrile (PAN) matrix infused with lithium ion sieves (LIS) H<SUB>1.6</SUB>Mn<SUB>1.6</SUB>O<SUB>4</SUB>. Characterization of the LIS/PAN NF confirmed its favorable structural and surface properties for effective Li<SUP>+</SUP> adsorption. The LIS/PAN NF was mechanically suitable as a microfiltration membrane with high water flux and low pressure requirement. Breakthrough experiments at varied feed concentrations (C<SUB>f</SUB>), seawater flowrates (F), and NF thicknesses (Z) revealed the dynamic adsorption behavior of the filter. The seawater residence time was most critical and must be kept ≥0.12min at any given C<SUB>f</SUB> and Z to maximize the Li<SUP>+</SUP> capacity of the filter. This can be conveniently achieved by adjusting the F of the process. Analogous to a packed bed system, the predictive power of nine breakthrough models were determined through non-linear regression analyses. Results reveal that bed-depth-space-time, Bohart-Adams (BA) and Thomas models adequately predicted the performance of the filter albeit BA exhibited the best agreement. Meanwhile, Wolborska failed to converge with any of the experimental results while Yoon-Nelson, Wang, Clark, dose-response, and modified dose-response were too simple to provide any meaningful information. Cycled Li<SUP>+</SUP> adsorption-desorption runs successfully collected and concentrated Li<SUP>+</SUP> in a mild acid stripping solution. After ten cycles, Li<SUP>+</SUP> was separated 155-1552 times more efficiently than Na<SUP>+</SUP>, K<SUP>+</SUP>, Mg<SUP>2+</SUP> and Ca<SUP>2+</SUP>. Overall results demonstrate the potential of LIS/PAN NF as an adsorbent membrane filter for continuous Li<SUP>+</SUP> mining from aqueous resources.