• International Journal of Stem Cells
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• 제16권2호
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• pp.234-243
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• 2023

The recent advances in human pluripotent stem cells (hPSCs) enable to precisely edit the desired bases in hPSCs to be used for the establishment of isogenic disease models and autologous ex vivo cell therapy. The knock-in approach based on the homologous directed repair with Cas9 endonuclease, causing DNA double-strand breaks (DSBs), produces not only insertion and deletion (indel) mutations but also deleterious large deletions. On the contrary, due to the lack of Cas9 endonuclease activity, base editors (BEs) such as adenine base editor (ABE) and cytosine base editor (CBE) allow precise base substitution by conjugated deaminase activity, free from DSB formation. Despite the limitation of BEs in transition substitution, precise base editing by BEs with no massive off-targets is suggested to be a prospective alternative in hPSCs for clinical applications. Considering the unique cellular characteristics of hPSCs, a few points should be considered. Herein, we describe an updated and optimized protocol for base editing in hPSCs. We also describe an improved methodology for CBE-based C to T substitutions, which are generally lower than A to G substitutions in hPSCs.

• 천문학회지
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• 제36권1호
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• pp.43-48
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• 2003

We introduce and describe performance of the 6-meter telescope of Seoul Radio Astronomy Observatory (SRAO). All the softwares and instruments except the antenna structure and its driving system are developed for ourselves. The SIS mixer type receiver resulted in the receiver noise temperature less than 50 K (DSB) over the whole 3-mm radio window. An autocorrelation spectrometer, developed first in Korea, provides maximum 50 MHz band width over 1024 channels. Antenna surface is measured and adjusted using template method and radio holography which resulted in a superb surface accuracy bet-ter than 30${\mu}m$. Accordingly, the aperture and beam efficiences amount to $70\%$ and $75\%$, respectively, largely independent of frequency in the 85 - 115 GHz range. It is also found that telescope pointing errors are less than 10" in both azimuth and elevation and that antenna gain is almost constant against elevation greater than $20^{\circ}$, without adjusting sub-reflector position. The SRAO 6-meter telescope is now fully operational and all these characteristics verify that observations are carried out with high precision and fidelity.

• 한국전자파학회논문지
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• 제15권8호
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• pp.737-743
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• 2004

120∼180 GHz 대역의 고정 튜닝방식을 사용한 SIS(Superconductor-Insulator-Superconductor) 접합 믹서를 개발 하였다. 이 믹서는 노베야마 전파천문대에서 제작된 6개 직렬 연결 Nb/Al-A1$_2$O$_3$/Nb SIS 접합 소자를 사용하였으며 석영유리 기판에 제작된 이 SIS 칩은 전 주파수 대역에서 입력신호 결합을 향상시키기 위해 half-height 도파관의 중심에 놓여 있다. 본 논문에서 개발된 SIS 믹서는 기계적인 튜닝 장치를 사용하지 않으며 RF 신호와 LO 전력은 냉각된 십자형 방향성 결합기를 통해서 믹서에 공급된다. 또한 IF 신호 손실을 줄이기 위해 SIS 믹서의 IF 출력 임피던스를 IF 증폭단의 50 $\Omega$ 입력 임피던스에 정합 시켰다. SIS 수신기의 측정된 DSB 잡음온도는 120∼180 GHz 대역에서 32∼131 K이며 본 논문에서 개발된 SIS 믹서는 대덕전파천문대의 14 m 전파망원경에 설치되어 전파천문 관측에 사용되고 있다.

• KSBB Journal
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• 제25권3호
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• pp.215-222
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• 2010

Monogenic disease의 치료를 위한 하나의 전략으로 viral vector를 이용한 gene therapy에 비해 독성이 적은 gene targeting 기술을 이용하기 위한 연구가 진행되고 있다. 이러한 연구의 주된 관점은 자연적인 HR의 낮은 효율을 개선하기 위한 DSB 유도 방법으로, 선택성을 높일 수 있는 긴 염기서열의 인식이 가능한 artificial endonuclease의 개발이다. 본 글에서는 이러한 artificial endonuclease 중, 가장 많이 연구 되고 있는 homing endonuclease와 zinc finger nuclease를 간략히 소개하였다. 전자와 후자 모두, 인식 서열에 대한 일정 수준의 tolerance (인식 서열 일부가 특이적이지 않아 다른 염기로 구성된 경우)가 존재하여, 일정한 비율로 다른 target을 절단할 수 있는 가능성이 존재한다. 이러한 점은, meganucleases를 치료 목적으로 이용할 때 세포 독성을 나타내는 근본원인 중 하나이다. 두 종 모두 이러한 특성을 가짐에도 불구하고, 완전한 비자연적인 후자보다는 전자의 경우가 보다 효과적이며 낮은 세포독성을 보이는 것으로 보고되고 있다. 물론 실험 조건이나 적용되는 세포 종류, 인위적인 단백질의 발현 정도에 따라 세포 독성유무 또는 정도에 차이가 나타남이 확인되고 있다. 이러한 사실들에 근거할 때, gene targeting을 유도하기 위한 artificial endonuclease의 서열 특이성을 증대시키는 것이 가장 중요하나, 그 외 여러 인자들에 대한 복합적인 연구 역시 필요함을 보여준다. 현재까지 실제 치료제로 쓰인 예는 없지만, 시험관내에서 보이는 결과와 모델 개체에서 이루어진 표적화정도, 관련된 단백질 치료제들이 지닌 잠재성을 비교할 때 매우 큰 가능성을 지니고 있음은 충분히 확인할 수 있다.

• Journal of Microbiology and Biotechnology
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• 제18권6호
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• pp.1186-1190
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• 2008

Single-chain Fv (scFv) antibody against c-Met is expected to be employed in clinical treatment or imaging of cancer cells owing to the important biological roles of c-Met in the proliferation of malignancies. Here, we show that the productivity of scFv against c-Met in Escherichia coli is significantly influenced by the orientation of its variable domains. We generated anti-c-Met scFv antibodies with two different domain orders (i.e., $V_L$-linker-$V_H$ and $V_H$-linker-$V_L$), expressed them in the cytoplasm of E. coli trx/gor deleted mutant, and compared their specific activities as well as their productivities. Productivity of total and functional anti-c-Met scFv with $V_H/V_L$ orientation was more than five times higher than that with $V_L/V_H$ format. Coexpression of DsbC enhanced the yield of soluble amounts of anti-c-Met scFv protein for both constructs. The purified scFv antibodies of the two different formats exhibited almost the same antigen-binding activities. We also compared the productivities and specific activities of anti-c-Met diabodies with $V_H/V_L$ or $V_L/V_H$ formats and obtained similar results to the case of scFv antibodies.

• 한국임상수의학회지
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• 제21권3호
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• pp.253-258
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• 2004

DNA double-strand break (DSB) is a serious treat for the cells including mutations, chromosome rearrangements, and even cell death if not repaired or misrepaired. Ku heterodimer regulatory DNA binding subunits (Ku70/Ku80) bound to double strand DNA breaks are able to interact with 470-kDa DNA-dependent protein kinase catalytic subunit (DNA-PKcs), and the interaction is essential for DNA-dependent protein kinase (DNA-PK) activity. The Ku80 mutants were designed to bind Ku70 but not DNA end binding activity and the peptides were treated in breast cancer cells for co-therapy strategy to see whether the targeted inhibition of DNA-dependent protein kinase (DNA-PK) activity sensitized breast cancer cells to ionizing irradiation or chemotherapy drug to develop a treatment of breast tumors by targeting proteins involved in damage-signaling pathway and/or DNA repair. We designed domains of Ku80 mutants, 26 residues of amino acids (HN-26) as a control peptide or 38 (HNI-38) residues of amino acids which contain domains of the membrane-translocation hydrophobic signal sequence and the nuclear localization sequence, but HNI-38 has additional twelve residues of peptide inhibitor region. We observed that the synthesized peptide (HNI-38) prevented DNA-PKcs from binding to Ku70/Ku80, resulting in inactivation of DNA-PK complex activity in breast cancer cells (MDA-465 and MDA-468). Consequently, the peptide treated cells exhibited poor to no DNA repair, and became highly sensitive to irradiation or chemotherapy drugs. The growth of breast cancer cells was also inhibited. These results demonstrate the possibility of synthetic peptide to apply breast cancer therapy to induce apoptosis of cancer cells.

• 통상정보연구
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• 제16권3호
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• pp.465-486
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• 2014

본 연구는 브라질 오렌지 주스사건의 제로잉에 대한 판결을 분석하고 있다. 미국은 동 사건에서 행정재심사에서 제로잉을 사용하였고, '거래에 대한 가중평균' 제로잉의 사용을 금한다는 이전 상소기구의 판례를 따르는 패널 판결이 내려졌다. 그러나 패널의 판결이 분쟁해결시스템내의 '안정성과 예측가능성'을 고려하여 나왔다는 점에 주목하면서, 동일한 법적 문제에 대한 상소기구의 판례에 대한 선례구속원칙이 암묵적으로 적용되고 있음을 판단할 수 있었다. 하지만 상소기구의 판결에 대한 패널의 이견 또한 확인할 수 있었다. 미국-오렌지주스사건의 판결이 오랫동안 이어져 온 제로잉 사용을 금지할 수 있을 것 같지만 제2.4조 하의 공정비교요건의 확대적용은 제로잉에 대한 쟁점을 지속시킬 가능성을 가지고 있는 것으로 판단된다. 다행스러운 것은 동 사건의 판결에 맞추어 행정재심사에 대한 제로잉을 금지하겠다는 미국의 발표가 있었다. 결국 제로잉에 대한 여러 분쟁이 남아있지만, 미국-오렌지주스 사건의 판결이 반덤핑협정하의 제로잉관행에 대한 최종적인 판결이라고 조심스럽게 예상해 본다.

• Journal of Forest and Environmental Science
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• 제34권3호
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• pp.199-208
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• 2018

Quercus floribunda (Tilonj oak) is among the five species of Quercus and an evergreen tree found in Kumaun Himalayan region. Timing of germination is a crucial event determining the success of seedling establishment and survival. The aims of the study were to investigate the effect of sowing date on the germination and morphological responses of Tilonj oak (Quercus floribunda), cultivated during the month of August-September, with supplementary irrigation. The experiment was conducted by sowing seeds at two dates with one month interval in glass house conditions at DSB Campus, Nainital, Uttarakhand, India. Present study revealed that higher germination percentage (46.67 %) was recorded at sowing time $S_2$ as compared to the sowing time $S_1$ (32.86 %). Germination percentage as well as seedling growth were affected by sowing date though the differences were insignificant. At both the sowing dates, highest germination percentage was recorded for large seeds (32.86-46.67 %) followed by medium (31.43-33.33 %) and lowest germination (6.67-7.14 %) was recorded for small size seeds. In all parameters assessed, sowing time $S_2$ had the best performance in combination with large seed size. Thus, September is suitable month for best germination, growth and seedling vigour and large seed size is recommended to silviculturists and tree planters. The information on seed germination and seedling growth is vital both for conservation and rehabilitation of degraded lands.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3431-3436
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• 2012

Objective: X-ray cross-complementing group 4 (XRCC4) is a major repair gene for DNA double-strand breaks (DSB) in the non-homologous end-joining (NHEJ) pathway. Several potentially functional polymorphisms of the XRCC4 gene have been implicated in breast cancer risk, but individually published studies showed inconclusive results. The aim of this meta-analysis was to investigate the association between XRCC4 polymorphisms and the risk of breast cancer. Methods: The MEDLINE, EMBASE, Web of science and CBM databases were searched for all relevant articles published up to June 20, 2012. Potential associations were assessed with comparisons of the total mutation rate (TMR), complete mutation rate (CMR) and partial mutation rate (PMR) in cases and controls. Statistical analyses were performed using RevMan 5.1.6 and STATA 12.0 software. Results: Five studies were included with a total of 5,165 breast cancer cases and 4,839 healthy controls. Meta-analysis results showed that mutations of rs2075686 (C>T) and rs6869366 (G>T) in the XRCC4 gene were associated with increased risk of breast cancer, while rs2075685 (G>T) and rs10057194 (A>G) might decrease the risk of breast cancer. However, rs1805377 (A>G), rs1056503 (G>T), rs28360317 (ins>del) and rs3734091 (A>G) polymorphisms of XRCC4 gene did not appear to have an influence on breast cancer susceptibility. Conclusion: Results from the current meta-analysis suggest that the rs2075685 (G>T) and rs6869366 (G>T) polymorphisms of the XRCC4 gene might increase the risk of breast cancer, whereas rs2075685 (G>T) and rs10057194 (A>G) might be protective factors.

• 천문학논총
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• 제27권3호
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• pp.71-80
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• 2012

We have developed superconducting mixer receivers for 129 GHz VLBI observation in Korean VLBI Network (KVN). The developed mixer has a radial waveguide probe with simple transmission line L-C transformer as a tuning circuit to its 5 series-connected junctions, which can have 125 - 165 GHz as the operation radio frequency (RF). For intermediate frequency (IF) signal path a high impedance quarter-wavelength line connects the probe to one end of symmetric RF chokes. The double side band (DSB) receiver noise of the mixer was about 40 K over 4 - 6 GHz IF band, whereas we achieved the uncorrected single side band (SSB) noise temperature of about 70 K and better than 10 dB image rejection ratio in 2SB configuration with 8 - 10 GHz IF band. Insert-type receiver cartridges employing the mixers have been under commission for KVN stations.