• Clinical and Experimental Pediatrics
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• 제51권1호
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• pp.54-61
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• 2008

목 적 : 특발성 신증후군의 연구를 위하여 병태생리 및 임상소견과 유사한 실험적 puromycin aminonucleoside(PAN) 신증을 이용하는데 본 연구는 배양한 사구체 상피세포에 대한 PAN의 영향을 통하여 신증후군의 주 원인 병태생리인 단백뇨의 기전을 밝히고자 하였다. 방 법 : 사구체 상피세포를 배양한 후 다양한 농도의 PAN과 항산화제를 투여하여 전자현미경관찰, 반응성 산소종 투과율 변화, confocal microscopy 등을 통하여 족세포성분의 변화를 관찰하였다. 결 과 : 사구체 상피세포의 초고배율소견에서 PAN에 의해 세포간극이 벌어지고 표면의 미세돌기가 단축되는 변화를 볼 수 있었다. 이러한 세포간극의 변화는 세포막부분의 ZO-1에 대한 면역형광검사에서도 확인할 수 있었다. DCF-DA로 측정한 반응성 산소종은 PAN에 의하여 농도에 따라 투여 2시간에 이미 유의한 증가를 보이나, 이러한 변화는 항산화제인 EGCG, probucol, vitamin C에 의해 감소하였다. 또한, 세포단층모델에서 투과율은 PAN에 의하여 농도에 따라 증가하나 항산화제에 의해 증가가 억제되었다. 세포골격구조인 ${\alpha}-actinin$은 사구체 상피세포의 세포질과 바깥 세포막부분으로 actin과 같이 분포하나 고농도의 PAN에 의해 세포질 바깥쪽의 일부분에 집중하는 형상으로 변하였다. 그러나 이러한 변화는 항산화제인 vitamin C의 처치에 의해 예방될 수 있었다. 세극막성분인 ZO-1는 고농도의 PAN에 의해 안쪽으로 이동하고 집중하는 형상으로 변하였으나, vitamin C의 처치에 의해 예방되었다. 이와 함께 ${\alpha}-actinin$과 ZO-1은 PAN에 의해 단백양이 감소하였으나 이는 항산화제에 의해 예방할 수 있었다. 결 론 : PAN은 사구체 상피세포의 반응성 산소종 생성을 증가시키고, 구조성분의 변화를 통하여 형태학적인 변화를 초래하며 이는 투과율의 증가로 나타났다. 이러한 변화들은 항산화제에 의해 어느 정도 억제할 수 있었음으로, PAN은 생체 외 사구체 상피세포에 산화스트레스기전을 통하여 구조적 변화와 이에 따른 단백뇨를 유발시키는 것으로 사료된다.

• Bulletin of the Korean Chemical Society
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• 제33권10호
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• pp.3407-3412
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• 2012

The Ti incorporation at Fe-site in the double perovskite lattice of $Sr_2FeNbO_6$ (SFNO) system is studied. The Ti concentration optimization yielded an efficient photocatalyst. At an optimum composition of Ti as x = 0.07 in $Sr_2Fe_{1-x}Ti_xNbO_6$, the photocatalyst exhibited 2 times the quantum yield for photolysis of $H_2O$ in presence of $CH_3OH$, than its undoped counterpart under visible light (${\lambda}{\geq}420nm$). Heavily Ti-doped $Sr_2Fe_{1-x}Ti_xNbO_6$ lattice exhibited poor photochemical properties due to the existence of constituent impurity phases as observed in the structural characterization, as well as deteriorated optical absorption. The higher electron-density acquired by n-type doping seem to be responsible for the more efficient charge separation in $Sr_2Fe_{1-x}Ti_xNbO_6$ (0.05 < x < 0.4) and thus consequently displays higher photocatalytic activity. The Ti incorporated structure also found to yield stable photocatalyst.

• 미생물학회지
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• 제33권1호
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• pp.1-6
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• 1997

5-Enolpyruvylshikimate 3-phosphate(EPSP) synthase는 방향족 아미노산을 생합성하는 shikimate phathway의 6번째 효소로 광범위 제초제인 glyphosate의 target enzyme이다. 본 연구에서는, glyphosate에 저해를 받지 않는 EPSP synthase를 개발하고자 하는 연구의 한 단계로서, 우선, EPSP synthase를 대량 발현시킬수 있는 expression vector인 pET-25b를 사용하여 발현시킨 다음, 발현된 효소가 periplasmic space로 transport되는지 또 발현된 단백질이 효소 활성을 가지고 있는지 확인하고자 하였다. 그 결과, pelB leader를 앞에 붙여 발현시킨 EPSP synthase는 periplasmic space로 제대로 transport되며, 단백질 생산 및 periplasmic space로의 수송은 induction 온도에 의해 크게 좌우된다는 것을 관찰하였다. Periplasmic space로 수송되는 EPSP synthase의 양은 $34^{\circ}C$에서 induction시켰을 때 가장 많은 것으로 나타났다. 한편, pET-25b를 이용하여 발현시킨 EPSP synthase는 C-terminal 부위에 HSV-tag, His-tag등 26개 아미노산이 더 있는 상태로 만들어지는데, His-tag은 $Ni^{2+}$-affinity chromatography를 통한 정제에, HSV-tag은 Western blotting을 통한 detection에 각각 이용할 수 있다. 또한, 이와 같이 발현된 recombinant EPSP synthase는 phosphocellulose resin에 결합하였다가 기질인 shikimate 3-phosphate와 phosphoenolpyruvate에 의해 elution되며, glyphosate에 의해 저해되는등 wildtype효소와 같은 효소 특성을 보였다.

• Tribology and Lubricants
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• 제30권3호
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• pp.146-155
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• 2014

In this study, we conduct fretting corrosion tests on tin-plated brass coupons to investigate the effect of temperature on fretting corrosion for various span amplitudes. We prepare a coupled fretting corrosion specimens using a tin-plated brass coupon with a thickness of $10{\mu}m$. One specimen is a flat coupon and the other specimen is a coupon with a protuberance in 1 mm radius, which is produced using 2 mm diameter steel ball. We conduct fretting corrosion tests at $25^{\circ}C$, $50^{\circ}C$, $75^{\circ}C$, $100^{\circ}C$ by rubbing the coupled coupons together at the contact between the flat and protuberance coupons. We measure electric resistance of the contact during the fretting corrosion test period. There is increase in resistance with fretting cycles. It is found that rate of increase in electric resistance becomes faster with increase in testing temperature. Magnitude of friction coefficient increases with fretting span amplitudes. And, change in friction coefficient becomes desensitized to the increment in span amplitude. Assuming that failure cycle is the cycle with an electric resistance of $0.01{\Omega}$, we find that failure lifetime ($N_f$) decreases with increase in testing temperature. Furthermore, based on the assumption that the damage rate of the connector is inversely related to the failure cycle, we calculate the activation energy for fretting damage to be 13.6 kJ/mole by using the Arrhenius equation. We propose a method to predict failure cycle at different temperatures for span amplitudes below $30{\mu}m$. Friction coefficients generally increase with increase in span amplitude and decrease in testing temperature.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.124-124
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• 2003

The successful development of embryos cloned by nuclear transfer (NT)have been dependent on a wide range of known factors including cell cycle of donor and recipient ooplast, oocyte quality, NT procedure and oocyte activation. The present study compared the development of cloned porcine embryos following different activation treatments. Cumulus-oocyte complexes (COCs) were aspirated from 26 mm follicles of slaughterhouse ovaries and cultured for 22 h in NCSU #23 medium supplemented with 10% porcine follicular fluid, 0.57 mM cysteine, 0.5 g/mL LH, 0.5 g/mL FSH and 10 ng/mL EGF. The COCs were further cultured for an additional 22 h in the same medium at $39{\cird}C$ in an atmosphere of 5% $CO_2$ in air, without hormonal supplements. Primary cultures of fibroblasts isolated from a female fetus on day 40 of gestation were established in DMEM + 15% FCS. For nuclear donation, cells at the 5th-6th passage were cultured in DMEM +0.5% FCS for 5 days in order to arrest the cells in G0/Gl. After enucleation, oocytes were reconstructed by transfer of donor cells and fusion with three DC pulses (1.4 KV/cm, 30 sec) in 0.28 M mannitol containing 0.01 mM $CaCl_2$ and 0.01 mM $MgCl_2$. Eggs were then divided into three treatment groups, control (without further treatment, Group 1), eggs cultured in 10 g/ml cycloheximide (CHX) for 5 h (Group 2), and eggs cultured in 1.9 mM 6-dimethylaminopurine (6-DMAP) for 5 h (Group 3). The eggs were then cultured in sets of 30 in 60 I drops of NCSU#23 supplemented with 4mg/ml BSA (essentially fatty acid free) until day 7 at $39{\circ}C$ in a humidified atmosphere of 5% $CO_2$. On day 4 the culture were fed by adding 20 I NCSU #23 supplemented with 10% FBS. Development rates into blastocysts were significantly higher (P<0.05) in Group 3 embryos compared to Group 1 controls ($27.6 \mu 2.7% vs. 20.1 \mu 4.1%$, respectively), but rates did not differ in Group 2 compared to control ($23.8 \mu 5.7%$). Total cell number in Group 3 blastocysts was however significantly higher (P<0.05) than in Groups 1 and 2 ($44.6 \mu 2.4 vs. 19.9 \mu 1.9 and 21.9 \mu 2.1$, respectively). These results suggest that 6-DMAP is more efficient than cycloheximide in the activation of electrically fused NT oocytes during in vitro production of cloned porcine embryos.

• Journal of Nutrition and Health
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• 제29권9호
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• pp.971-980
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• 1996

The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

• KSBB Journal
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• 제8권2호
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• pp.97-103
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• 1993

세포질 유전형질의 도입을 성공적으로 수행코자 세질 웅성 불임(eMS; cytoplasmic male sterili­t ty) 계통의 담배 MS(male sterility) Burley 21의 callus tissue에서 cytoplast를 분리시키고 이와 Ni­c cotiana tabacum Burley 64의 mesophyll protoplast 를 PEG방법에 의한 세포융합으로 cybrid cell올 유도할 수 있었다. cytoplast의 분라는 density gradient solution의 osmolarity를 일정하게 하는 것보다 gradient간의 차이가 더 효과적이였으며 분리된 cytoplast의 양은 재료로 이용된 protoplast의 20-30% 정도를 나타 내었다. 또한 cytoplast와 protoplast의 융합은 P PEG농도 50%에셔 효과적이었다.

• Fisheries and Aquatic Sciences
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• 제13권1호
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• pp.36-48
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• 2010

An agar-degrading Agarivorans sp. AG17 strain was isolated from the red seaweed Grateloupia filicina collected from Jeju Island. A beta-agarase gene from Agarivorans sp. AG17 was cloned and designated as agrA. agrA has a 2,985 bp coding region encoding 995 amino acids and was classified into the glycoside hydrolase family (GHF)-50. Predicted molecular mass of the mature protein was 105 kDa. His-tagged agrA was overexpressed in Escherichia coli and purified as a fusion protein. The enzyme showed 158.8 unit/mg specific activity (optimum temperature at $65^{\circ}C$ and pH 5.5 in acetate buffer) with unique biochemical properties (high thermal and pH stabilities). Enzyme produced neoagarohexaose, neoagarotetraose and neoagarobiose by degrading agar, and hydrolyzed neoagaro-oligosaccharides were biologically active. Hence the purified enzyme has potential for use in industrial applications such as the development of cosmetics and pharmaceuticals.

• 혜화의학회지
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• 제8권1호
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• pp.115-129
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• 1999

To evaluate radioprotective effects of Kamisagoonjatang(KST), Kamijihwangtang (KJT) and Kamigoonjajihwangtang(KKJT), studies were done experimentally. The results were obtained as follows: 1. By FACS analysis after exposure to radiation by Liniac, T cell and T-helper cell were significantly increased in KST treated group and also B cell and macrophage in KJT treated group while splenocytes were significantly decreased in control group. 2. WBC, PLT were significantly increased in KKJT treated group as compared with control group after exposure to radiation by Liniac. 3. In histological changes of jejunum of $BALB{\backslash}C$ mice after after exposure to radiation by Liniac, exclusion and fusion of villi were decreased in all groups as compared with control group. 4. In the observation of morphological changes by SEM and TEM after radiation by Liniac, KKJT, KJT and KST inhibited demage of internal structures such as mitochondria, ESR and golgi of jejunum cells in order as compared with control group. From above results it was concluded that KJT, KST and KKJT could be usefully applied for protection from damage by radiotherapy to cancer.

• Applied Microscopy
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• 제25권2호
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• pp.20-28
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• 1995

This study observes the change of small intestine mucosa paneth cell by changing the amount of radiation to rat. It uses the rat(Wistar) of 250-300g as the experimental animal and irradiation equipment is Gammacell 3000Elan System. and the irradiation is conducted for 500Rad group for 34sec., 1000rad for 68sec., and 1500Rad for 102sec. once on the whole body of each group, eachgroup is anesthetized with ether after 24hours. its small intestine is extrated and then it is observed by transmission electronic microscopy. The experimental results are as follows : 1. 500 Rad Group The Slightly elongated form of mitochondria and rough endoplasmic reticulum are observed in 500 Rad group. 2. 1000 Rad Group Golgi apparatus is appeared as the extended plasmodium, secretory granules exist only external membrane due to the self-fusion, the number of mitochondria that are changed as L-type are reduced, rough endoplasmic reticulum is distributed with the expanded form. 3. 1500 Rad Group The number of Golgi apparatus and granules is remarkably reduced, mitochondria is changed into C-type and free ribosomes can be observed instead of the reduction of rough endoplasmic reticulum.