Ti-Al-Fe계 합금의 고온산화거동에 미치는 Fe의 영향

윤장원,현용택,김정한,염종택,윤석영,Yoon, Jang-Won,Hyun, Yong-Taek,Kim, Jeoung-Han,Yeom, Jong-Taek,Yoon, Seog-Young 한국재료학회 2011 한국재료학회지 Vol.21 No.7

In this paper, high temperature oxidation behavior of newly developed alloys, Ti-6Al-4Fe and Ti-6Al-1Fe, is examined. To understand the effect of Fe on the air oxidation behavior of the Ti-Al-Fe alloy system, thermal oxidation tests are carried out at $700^{\circ}C$ and $800^{\circ}C$ for 96 hours. Ti-6Al-4V alloy is also prepared and tested under the same conditions for comparison with the developed alloys. The oxidation resistance of the Ti-Al-Fe alloy system is superior to that of Ti-6Al-4V alloy. Ti-6Al-4V shows the worst oxidation resistance for all test conditions. This is not a result of the addition of Fe, but rather it is due to the elimination of V, which has deleterious effects on high temperature oxidation. The oxidation of the Ti-Al-Fe alloy system follows the parabolic rate law. At $700^{\circ}C$, Fe addition does not have a noticeable influence on the amount of weight gain of all specimens. However, at $800^{\circ}C$, Ti-6Al-4Fe alloy shows remarkable degradation compared to Ti-6Al-1Fe and Ti-6Al. It is discovered that the formation of $Al_2O_3$, a diffusion resistance layer, is remarkably hindered by a relative decrease of the ${\alpha}$ volume fraction. This is because Fe addition increases the volume fraction of ${\beta}$ phase within the Ti-6Al-xFe alloy system. Activities of Al, Ti, and Fe with respect to the formation of oxide layers are calculated and analyzed to explore the oxidation mechanism.

돼지 체세포 복제 수정란의 자가 사멸 분석

류지은,윤종택,Ryu, Ji-Eun,Yoon, Jong-Taek 한국수정란이식학회 2018 한국동물생명공학회지 Vol.33 No.3

The purpose of this study is to examined the electrofusion and activation conditions for the production of porcine somatic cell nuclear transfer (SCNT) embryos. In this study, immature oocytes were cultured in TCM-199 with and without hormones for 22 hours. Skin fibroblasts cells of porcine were transferred into the perivitelline space of enucleated in vitro matured oocytes. Cell fusion was performed with two different pulses that each one pulse (DC) of 1.1 kV/cm or 1.5 kV/cm for $30{\mu}sec$. After fusion subsequent activation were divided into three groups; non-treatment (control) and treatment with 2 mM 6-DMAP or $7.5{\mu}g/ml$ cytochalasin B for 4 hours. Transferred embryos were cultured in PZM-3 (Porcine Zygote Medium-3) in $5%\;CO_2$ and 95% air at $39^{\circ}C$ for 7 day. Apoptosis-related genes (Caspase-3, BCL-2, mTOR, and MMP-2) were analyzed by immunofluorescence staining. There was no significant difference between two different electrofusion stimuli in the cleavage rate; $64.9{\pm}4.8%$ in 1.1 kV/cm and $62.7{\pm}4.0%$ in 1.5 kV/cm. However, blastocyst formation rate (%) was significantly different among three different activation groups (no treatment, 2 mM 6-DMAP or $7.5{\mu}g/ml$ cytochalasin B) combined with electrofusion of 1.1 kV/cm. The blastocyst formation rate was $12.6{\pm}2.5$, $20.0{\pm}5.0$, and $34.9{\pm}4.3%$ in control, 2 mM 6-DMAP, and $7.5{\mu}g/ml$ cytochalasin B, respectively. Immunofluorescence data showed that expression levels of caspase-3 in SCNT embryos undeveloped to blastocyst stage were higher than those in the blastocyst stage embryos. Expression levels of Bcl-2 in blastocyst stage embryos were higher than those in the arrested SCNT embryos. These results showed that the combination of an electric pulse (1.1 kV/cm for $30{\mu}sec$) and $7.5{\mu}g/ml$ cytochalasin B treatment was effective for production of the porcine SCNT embryos.

소 난자의 체외성숙시 난구세포와 난세포질에서 다르게 발현되는 Matrix Metalloproteinases의 분석

김상환,윤종택,Kim, Sang-Hwan,Yoon, Jong-Taek 한국수정란이식학회 2018 한국동물생명공학회지 Vol.33 No.3

To determine the differences in the in-vitro ovum maturation process of bovine, we compared the expression of MMPs in these oocytes and cumulus cell throughout oocytes maturated. In an attempt to investigate the effect of MMP activation and inhibitors in total protein of cumulus cell and, oocytes during oocytes maturation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9), TIMPs (TIMP-2 and TIMP-3), as well as their expression profiles (Real-time PCR, Gelatin Zymography and ELISA). Our results that the bovine oocytes MMP-2 and MMP-9 level was significantly associated with the rate of maturity of oocytes (P<0.05). In cumulus cell, MMP-2 was highly expressed in all stages of the oocyte's maturation. The final oocytes maturation exhibited strong gelatinase activity. There was no significant correlation between cumulus cell MMP-9 and the maturation rate of oocytes. However, for the oocyte cytoplasm MMP-9 expression was significant correlation to the maturation oocytes. There was no significant correlation between cumulonimbus cells MMP-9 and oocyte maturation rates; however, for oocyte cytoplasm, MMP-9 expression was significantly correlated with mature oocyte. However, the TIMP-1 and TIMP-2 protein expression patterns are not correlated with the maturation rate of the oocyte. Our results suggest that MMP different expression pattern may regulate the morphological remodeling of oocyte's in the cumulus cell. Further, the MMP-2 expression has a strong relation with a higher maturation rate of the oocyte.

소의 초기 임신 황체에서 PAPP-A와 $20{\alpha}$-HSD의 발현 양상

김대승,김상환,윤종택,Kim, Dae-Seung,Kim, Sang-Hwan,Yoon, Jong-Taek 한국수정란이식학회 2011 한국동물생명공학회지 Vol.26 No.1

This study was performed to the expressions of pregnancy-associated plasma protein-A (PAPP-A) and 20alpha-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) in bovine corpus luteum during early pregnancy. To determine the function of PAPP-A gene during early pregnancy, we collected corpus luteum samples on 30, 60 and 90 days of pregnancy in bovine. The mRNA expression of PAPP-A, $20{\alpha}$-HSD, progesterone-receptor (PR) and insulin-like growth factor binding protein4 (IGFBP4) gene was conducted by Real-time PCR. In parallel with mRNA levels, The protein expressions of PAPP-A and $20{\alpha}$-HSD were detected by immunological analysis. The mRNA expressions $20{\alpha}$-HSD and PAPP-A significantly increased on day 90 in the corpus luteum during pregnancy. The mRNA expression of PR and JGFBP4 in the corpus luteum progressively was enhanced at 30 to 60 day, but decreased on 90 day of pregnancy in the corpus luteum. The expression patterns of these genes, PAPP-A and $20{\alpha}$-HSD were similar pattern in these tissues. In conclusion, PAPP-A and $20{\alpha}$-HSD activity in corpus luteum could be played a role for early pregnancy manifestation.

칡소 모색별 피부세포 배양 시 Estrogen 과 Testosterone 첨가가 Melanogenesis 관련 유전자의 발현에 미치는 영향

서희경,이지혜,김상환,이호준,윤종택,Seo, Hui-Gyeong,Lee, Ji-Hye,Kim, Sang-Hwan,Lee, Ho-Jun,Yoon, Jong-Taek 한국수정란이식학회 2018 한국동물생명공학회지 Vol.33 No.3

The purpose of this study is to expression pattern of melanogenesis associate genes on cultured melanocyte layer cells in Korean Brindle Cattle(Dark, Brindle and Yellow) were analyzed to evaluate the effects of sex hormones on the control of melanogenesis pathways. Korean Brindle Cattle(Dark, Brindle and Yellow) melanocyte in the skin cells was collected. after the addition of estrogen and testosterone, the culture was analyzed for expression of cell activity and melanin genes for 72 hours. For the analysis of estrogen in different coat color other than the melanogenesis-related genes it is increasingly yellow showed low expression. in particular, the cells of the brindle coat color is low active and expression of genes. However, the testosterone was low, the expression of cell activity inhibiting MMP-2. the expression of melanin genes actually showed a tendency to increase gradually, which is testosterone compared with the estrogen to be considered that affect the skin cell layer brindle coat color. In this study, stimulation with estrogen triggered the inhibition of MC1R of the melanocyte in brindle coat color, but testosterone is induced MC1R in melanocyte. Therefore, considered the eumelanin or phaeomelanin activation are controlled caused by differential expression of sex hormones on melanocyte in Korean Brindle Cattle.

체세포 복제란 이식우의 분만 전.후 TGF-${\beta}_1$ 단백질 농도

황성수,장유민,고응규,양병철,임기순,김명직,민관식,윤종택,김창근,성환후,Hwang, Seong-Soo,Chang, Yoo-Min,Ko, Yeoung-Gyu,Yang, Byong-Chul,Im, Gi-Sun,Kim, Myong-Jik,Min, Kwan-Sik,Yoon, Jong-Taek,Kim, Chang-Keun,Seong, Hwan-Hoo 한국동물번식학회 2008 Reproductive & developmental biology Vol.32 No.1

본 연구는 체세포 복제란 이식우의 분만에 있어서 혈중 스테로이드호르몬, TGF-${\beta}_1$ 농도와 분만지연의 상관 관계를 살펴보고자 실시하였다. 대조군으로는 인공수정(AI)을 통하여 임신한 암소(cow)들을 사용하였다(AI-R). 모든 AI-R들은 자연분만(n=5, 임신 $284{\pm}0.71$일)을 하였다. 분만징후를 보이지 않는 체세포 복제란 이식우(n=5, SCNT-R)들은 분만 예정일보다 10일 정도 지난 임신 292일째에 제왕절개(Caesarean section, C-sec)를 실시하여 분만하였다. 혈액 및 태반 샘플을 분만 전.후에 채취하여 형태 및 중량 등을 측정하였다. 혈장호르몬인 Progesterone(P4)와 Estradiol-$17{\beta}$(E2) 농도는 방사선동위원소 면역 분석 시험(RIA) 방법을 이용하여 측정하였다. 혈장 및 태반분엽의 TGF-${\beta}_1$ 농도는 ELISA 방법으로 측정하였다. SCNT-R에서 회수한 태반의 무게는 AI-R의 것과 비교하여 유의적으로 무거웠다(p<0.05). 분만 직전 SCNT-R들의 혈장 내 P4 농도는 AI-R들의 그것과 비교하여 유의하게 높았다(p<0.01). 하지만 SCNT-R들의 혈장 내 E2 농도는 AI-R과 비교하여 상대적으로 낮게 나타났다(p<0.01). 한편, 분만 전.후 SCNT-R들에서 혈장 또는 태반분엽의 TGF-${\beta}_1$ 단백질 발현 수준은 AI-R들과 비교하여 각각 유의적으로 높은 수준을 유지하였다(p<0.01). 이상의 결과를 종합하여 보면, 분만 시 P4 및 E2의 이상 발현과 높은 수준의 혈장 및 태반 내 TGF-${\beta}_1$ 단백질은 체세포 복제태아의 분만지연을 야기하는 중요한 요인들 중의 하나일 것이라 사료된다. This study was performed to investigate the correlations between steroids and TGF-${\beta}_1$ levels and delayed parturition in SCNT clone calving. The recipients pregnant by AI were used as control (AI-R). All AI-R were labored by natural delivery (n=5, day $284{\pm}0.71$ of pregnancy). The recipients pregnant by SCNT embryo (SCNT-R) showing no signs of delivery about 10 days after expected date were operated by Caesarean section (n=5, day 292). The blood and placentome samples were obtained and weighed at parturition. The concentrations of plasma progesterone (P4) and Estradiol-$17{\beta}$ (E2) were measured by radioimmunoassay (RIA). The levels of plasma and placental TGF-${\beta}_1$ levels were examined by ELISA. The placentomes from SCNT-R were overweight (p<0.05) compared to those of AI-R. The plasma P4 (p<0.01) level in SCNT-R at parturition was significantly higher compared to that of AI-R. In contrast, the plasma E2 level in the SCNT-R was significantly lower compared to that of AI-R (p<0.05). The plasma and placental TGF-${\beta}_1$ protein levels in the SCNT-R were significantly higher than those of AI-R at parturition, respectively (p<0.01). Based on these results, aberrant expressions of steroid hormones and high levels of plasma and placental TGF-${\beta}_1$ protein at parturition may be one of the key indicators on delayed parturition of SCNT clone calving.

소의 체세포핵이식태반과 정상태반간의 차등 발현 유전자 분석

유성란 ( Seong Lan Yu ),정행진 ( Hang Jin Jeong ),상병찬 ( Byung Chan Sang ),류승희 ( Seung Heui Ryoo ),정기철 ( Kie Chul Jung ),윤종택 ( Jong Taek Yoon ),성환후 ( Hwan Hoo Seong ),진동일 ( Dong Il Jin ),이준헌 ( Jun Heon Lee ) 한국동물자원과학회 2008 한국축산학회지 Vol.50 No.5

장기이식과 체세포복제 돼지의 이용

윤종택 中央大學校 遺傳工學硏究所 2001 遺傳工學硏究論集 Vol.14 No.1

Organ transplantation is still very rarely performed. Transplantation is clearly established as the treatment of choice for many patients suffering from end-stage organ failure, as major advances in techniques and the development of new immunosupressive therapics have significantly improved the outcome of clinical transplantation during the past two decades. Attention has been focused on the potential use of animal organs as an alternative and promising solution to this serious problem. Recently, the modification of pigs, which are the only animals now being considered as a source of source organs for human transplantation, have been pursued by means of genetic engineering techniques.

血淸 및 호르몬 添加가 소 卵胞卵의 體外成熟에 미치는 影響

尹鐘澤,鄭英彩,金昌根 中央大學校 遺傳工學硏究所 1991 遺傳工學硏究論集 Vol.4 No.1

These studies were carried out to investigate the effects of sera and hormones added to maturation medium on the in vitro maturation(IVM) in bovine follicular oocytes. The results obtained were as follows: 1. The IVM rate of oocytes was increased by addition of 10 to 15% FCS to m-KRB and DM and 20% FCS to Ham's F10, respectively. The addition of ECS to TCM199 was superior to that of FCS, but no difference in the IVM rate between ECS and FCS added to m-KRB. 2. The IVM rate in the media added with hormones was increased and the rate in m-KRB was higher when FSH was added to medium instead of HCG. The highest IVM rate was obtained in the TCM199 added with 10% ECS and LH. 3. The IVM rate of oocytes in the media added with sera was higher than that of hormones. 4. In conclusion, the present studies indicate that the addition of sera and hormones can improved the in vitro maturation of bovine follicular oocytes.

소난포난의 체외발생에 있어서 혈청첨가와 과립막세포 및 난관상피 세포와의 공배양 효과

정광조,정영채,윤종택,김창근 中央大學校 遺傳工學硏究所 1992 遺傳工學硏究論集 Vol.5 No.1

본 연구는 미성숙난포란을 체외성숙 후 체외수정하여 cell block stage를 극복하고 배를 발생시켜 수정란이식에 의하여 송아지를 생산할수 있는 체외수정 체계를 확립하기위하여 도살장에서 도살되는 한우와 홀스타인 소로부터 체취된 난포란을 혈청과 과립막세포와 체외성숙후 정자처리 방법에 따라 체외수정을 실시하여 과립막세포 및 난관상피세포와 체외배양하여 초기배 발생능을 조사한바 다음과 같은 결과를 얻었다. 1.TCM199 성숙배양액에 FCS와 ECS10%만을 첨가하였을 때 난포란의 체외성숙율은 66.6%와 72.7%이었으나 FCS와 ECS10%를 첨가하고 과립막세포와 공배양하였을 때는 81.3%와 84.0%로 체외성숙율이 향상되었다. 2.TCM 성숙배양액에 10% FCS만을 첨가하고 난포란을 체외성숙시킨 후 caffeine 또는 caffeine-heparin 처리된 정자와 체외수정하였을 때 난할율은 19.0%와 18.4%이었다. 또한 FCS10%와 과립막세포와 공배양하여 체외성숙시킨 난포란의 난할율은 25.0%와 22.9%이었다. 3.TCM199성숙배양액에 10% FCS만을 첨가하여 성숙시킨 후 caffeine-heparin 처리된 정자와 체외수정하여 TCM199성숙배양액에 10% FCS만을 첨가하여 체외배양하였을 때 난포란의 발생율은 22.2%이었으나 상실배까지는 발생되지 않았다. 4.10% FCS 및 과립막세포와 공배양하여 체외성숙 시킨 후 체외수정한 다음 난관상피세포와 공배양하여 체외배양하였을 때 배발생율은 46.0%이었고 과립막세포와 체외배양하였을 때는 50.2%이었다. 5.TCM199 성숙배양액에 10% ECS만을 첨가하여 과립막세포와 공배양하여 체외수정한다음 난관상피세포와 공배양하여 체외배양 하였을 때 배발생율은 45.2%이었고 과립막세포와 체외배양하였을 때는 51.4%이었다. 6.TCM199 성숙배양액에 10% FCS및 과립막세포와 체외성숙시킨 난포란을 caffeine처리된 정자와 체외수정하여 난관상피세포와 공배양하여 체외배양하였을 때 배발생율은 38.4%이었다. Caffeine-heparin 처리된 정자와 체외수정하여 난관상피세포와 공배양하여 체외 배양하였을 때 배발생율은 42.6%이었고 과립막세포와 체외배양하였을 때 52.2%이었다. 7.10% ECS 및 과립막세포와 공배양하여 체외성숙시킨 한우 난포란의 배발생율은 난관상피세포와 공배양하여 체외배양하였을 때 41.8%이었고 과립막세포와 체외배양하였을 때는 57.1%이었다. 한편 FCS10% 및 과립막세포와 공배양하여 체외성숙시킨 후 난관상피세포와 공배양하여 체외배양하였을 때 배발생율은 42.0%이었고 과립막세포와 체외배양하였을 때는 48.4%이었다. These studies were carried out to investigate the effects of sera(fetal calf serun: FCS: and estrus cow serum: ECS) and granulosa cells(GC) added to maturation medium for in vitro maturation and developmental capacity of bovine follicular oocytes co-cultured with bovine epithelial cells(BOED) and granulosa cells in vitro. The ovaries were obtained from slaughtered Korean native cows and Holsteines, The follicular oocytes were cultured in TCM199 medium with sera and granulosa cells for 24hrs. The matured oocytes were fertilized in vitro using frozen-thawed sprematozoa in BO solution containing caffeine(5mM/ml) or caffeine(5mM/ml)-heparin(10mg/ml), and then the oocytes were co-cultured with bovine epithelial cells or granulosa cells. The results obtained were summarized as follows: 1.In vitro maturation rates of bovine follicular oocytes cultured in TCM199 with 10% FCS or ECS were 66.6% and 72.2%, but the case of addition of 10% FCS or ECS to TCM199 co-cultured with granulosa cells were 81.3% and 84.0%, respectively. IVM rate of these TCM199 added to granulosa cells was higher than that of media without granulosa cells. 2.Cleavage rates of follicular oocytes matured in TCM199 with 10% FCS alone and then fertilized in vitro by sperm treated with caffeine or caffeine-heparin were 19.0% and 18.4%, but the case of addition of 10% FCS to TCM199 with granulosa cells were 25.0% and 22.9%. 3.Cleavage rates of bovine oocytes cultured with 10% alone and fertilized in vitro by sperm treated with caffeine-heparin was 22.2%, but there was no blastocyst. 4.When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC, embryo developments of bovine oocytes co-cultured with BOEC of GC were 46.0% and 50.2%, respectively. 5.When bovine follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6.When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC and then fertilized in vitro by sperm treated with caffeine, embryo developments of bovine oocytes co-cultured with BOEC were 38.4% and 51.4%, respectively. But those of bovine oocytes co-cultured with GC were 52.2% by sperm treated with caffeine-heparin. 7.When Korean native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developments of the bovine oocytes co-cultured with BOEC and GC were 41.8% and 57.1%. But with FCS 10% those of the bovine oocytes co-cultured with BOEC and GC were 42.0% and 48.4%, respectively. 8.When Holsteine's follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments of the bovine oocytes co-cultured with BOEC and GC were 50.0% and 56.9%, but with ECS 10% those of the bovine oocytes co-cultured with BOEC and GC were 52.2% and 56.5%, respectively. 9.The Present results indicate that sera and GC added to TCM199 are the effective components for in vitro maturation of bovine oocytes and for in vitro development, developmental capacity of bovine oocytes is improved when they are co-cultured with the somatic cells such as BOEC and GC.