• BMB Reports
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• 제46권3호
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• pp.163-168
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• 2013

The AML1 gene is an essential transcription factor regulating the differentiation of hematopoietic stem cells into mature blood cells. Though at least 12 different alternatively spliced AML1 mRNAs are generated, three splice variants (AML1a, AML1b and AML1c) have been characterized. Here, using the reverse transcription-polymerase chain reaction with outward-facing primers, we identified a novel non-polyadenylated transcript from the AML1 gene, with exons 5 and 6 scrambled. The novel transcript resisted RNase R digestion, indicating it is a circular RNA structure that may originate from products of mRNA alternative splicing. The expression of the novel transcript in different cells or cell lines of human and a number of other species matched those of the canonical transcripts. The discovery provides additional evidence that circular RNA could stably exist in vivo in human, and may also help to understand the mechanism of the regulation of the AML1 gene transcription.

• Molecules and Cells
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• 제41권8호
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• pp.733-741
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• 2018

Mutations in spliceosome components have been implicated in carcinogenesis of various types of cancer. One of the most frequently found is U2AF1 S34F missense mutation. Functional analyses of this mutation have been largely limited to hematological malignancies although the mutation is also frequently seen in other cancer types including lung adenocarcinoma (LUAD). We examined the impact of knockdown (KD) of wild type (wt) U2AF1 and ectopic expression of two splice variant S34F mutant proteins in terms of alternative splicing (AS) pattern and cell cycle progression in A549 lung cancer cells. We demonstrate that induction of distinct AS events and disruption of mitosis at distinct sub-stages result from KD and ectopic expression of the mutant proteins. Importantly, when compared with the splicing pattern seen in LUAD patients with U2AF1 S34F mutation, ectopic expression of S34F mutants but not KD was shown to result in common AS events in several genes involved in cell cycle progression. Our study thus points to an active role of U2AF1 S34F mutant protein in inducing cell cycle dysregulation and mitotic stress. In addition, alternatively spliced genes which we describe here may represent novel potential markers of lung cancer development.

• 한국발생생물학회지:발생과생식
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• 제22권1호
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• pp.73-83
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• 2018

This study investigates the endoplasmic reticulum (ER) stress and subsequent apoptosis in duced during somatic cell nuclear transfer (SCNT) process of porcine SCNT embryos. Porcine SCNT and in vitro fertilization (IVF) embryos were sampled at 3 h and 20 h after SCNT or IVF and at the blastocyst stage for mRNA extraction. The x-box binding protein 1 (Xbp1) mRNA and the expressions of ER stress-associated genes were confirmed by RT-PCR or RT-qPCR. Apoptotic gene expression was analyzed by RT-PCR. Before commencing SCNT, somatic cells treated with tunicamycin (TM), an ER stress inducer, confirmed the splicing of Xbp1 mRNA and increased expressions of ER stress-associated genes. In all the embryonic stages, the SCNT embryos, when compared with the IVF embryos, showed slightly increased expression of spliced Xbp1 (Xbp1s) mRNA and significantly increased expression of ER stress-associated genes (p<0.05). In all stages, apoptotic gene expression was slightly higher in the SCNT embryos, but not significantly different from that of the IVF embryos except for the Bax/Bcl2L1 ratio in the 1-cell stage (p<0.05). The result of this study indicates that excessive ER stress can be induced by the SCNT process, which induce apoptosis of SCNT embryos.

• 생명과학회지
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• 제7권3호
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• pp.161-166
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• 1997

HIV-1 Rev단백질은 바이러스의 구조단백질의 발현조절에 매우 중요한 역할을 하며RNA가 1차발현된 원형대로 또는 일부만이 절단/재조합된 상태로 세포질에 집적되는 것을 가능하게 한다. env gene에 존재하는 Rev-responsive element RNA는 복잡한 RNA구조를 지니면서 Rev의 기능을 위하여 필수적으로 필요시 된다. 그러나 이러한 절차의 완전한 진행을 위해서는 핵단백질이 요구되는 것으로 추정된다. 본 연구에서는 electrophoretic mobility shift, UV-crosslinking 또는 SDS/PAGE등의 실험을 통하여 36/37, 56, 41, 76, 150 kD등의 핵단백질등이 RRE RNA와 결합반응하는 것을 발견하였으며 특히 36/37 과 56 kD 단백질을 5분간의 자외선조사에 의해 특히 RRE RNA에 결합하는 핵단백질들은 gel mobility shift assay에서 Rev RNA인식결합에 경쟁적인 특징을 나타내고 있다.

• Earthquakes and Structures
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• 제17권3호
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• pp.297-306
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• 2019

The extension of existing RC buildings is a challenging process, which requires efficient connection between existing and new materials to guarantee load transferring between the lap-spliced longitudinal columns' reinforcement. Therefore, the length of the columns' starter bars is a crucial factor, which decisively affects the seismic response of the new columns. In particular, when the length of the starter bars is short, then the length of the lap splices of reinforcement is inadequate to ensure load transfer between steel bars and concrete, with an indisputable detrimental impact on the seismic behaviour of the columns. Moreover, in most of the existing RC buildings the column starter bars are of particularly short length, while they have probably been bent, cut or corroded. In the present study, the effectiveness of both welded rebar and mechanical splices of reinforcement in ensuring load transferring between the starter bars and the longitudinal reinforcement of the new column was experimentally evaluated. Four cantilever column subassemblages were constructed and subjected to earthquake-type loading. Three of the specimens were used to examine different types of shielded metal arc welding (SMAW), while in the fourth subassemblage mechanical splices were tested. The hysteretic response of the columns was evaluated and compared to the behaviour of a fifth specimen with continuous reinforcement, tested by Kalogeropoulos and Tsonos (2019). Test results clearly demonstrated that the examined types of SMAW were equally satisfactory in ensuring the ductile seismic performance of the columns, while the mechanical splices found to be more susceptible to exhibit slipping of the bars.

• Biomolecules & Therapeutics
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• 제30권3호
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• pp.265-273
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• 2022

Resistance to chemotherapeutic drugs is a significant problem in the treatment of colorectal cancer, resulting in low response rates and decreased survival. Recent studies have shown that shikonin, a naphthoquinone derivative, promotes apoptosis in colon cancer cells and cisplatin-resistant ovarian cells, raising the possibility that this compound may be effective in drug-resistant colorectal cancer. The aim of this study was to characterize the molecular mechanisms underpinning shikonin-induced apoptosis, with a focus on endoplasmic reticulum (ER) stress, in a 5-fluorouracil-resistant colorectal cancer cell line, SNU-C5/5-FUR. Our results showed that shikonin significantly increased the proportion of sub-G₁ cells and DNA fragmentation and that shikonin-induced apoptosis is mediated by mitochondrial Ca²⁺ accumulation. Shikonin treatment also increased the expression of ER-related proteins, such as glucose regulatory protein 78 (GRP78), phospho-protein kinase RNA-like ER kinase (PERK), phospho-eukaryotic initiation factor 2 (eIF2α), phospho-phosphoinositol-requiring protein-1 (IRE1), spliced X-box-binding protein-1 (XBP-1), cleaved caspase-12, and C/EBP-homologous protein (CHOP). In addition, siRNA-mediated knockdown of CHOP attenuated shikonin-induced apoptosis, as did the ER stress inhibitor TUDCA. These data suggest that ER stress is a key factor mediating the cytotoxic effect of shikonin in SNU-C5/5-FUR cells. Our findings provide an evidence for a mechanism in which ER stress leads to apoptosis in shikonin-treated SNU-C5/5-FUR cells. Our study provides evidence to support further investigations on shikonin as a therapeutic option for 5-fluorouracil-resistant colorectal cancer.

• Animal Bioscience
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• 제35권1호
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• pp.22-28
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• 2022

Objective: Endoplasmic reticulum (ER) stress engages the unfolded protein response (UPR) that serves as an important mechanism for modulating hepatic fatty acid oxidation and lipogenesis. Chronic fasting in mice induced the UPR activation to regulate lipid metabolism. However, there is no direct evidence of whether negative energy balance (NEB) induces ER stress in the liver of cows. This study aimed to elucidate the relationship between the NEB attributed to feed deprivation and ER stress in bovine hepatocytes. Methods: Blood samples and liver biopsy tissues were collected from 6 non-lactating cows before and after their starvation for 48 h. The blood non-esterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA) and glucose level were analyzed. Real-time quantitative polymerase chain reaction and Western blotting were used to explore the regulation of genes associated with UPR and lipid metabolism. Results: The starvation increased the plasma BHBA and NEFA levels and decreased the glucose level. Additionally, the starvation caused significant increases in the mRNA expression level of spliced X-box binding protein 1 (XBP1s) and the protein level of phosphorylated inositol-requiring kinase 1 alpha (p-IRE1α; an upstream protein of XBP1) in the liver. The mRNA expression levels of peroxisome proliferator-activated receptor alpha and its target fatty acid oxidation- and ketogenesis-related genes were significantly upregulated by the starvation-mediated NEB. Furthermore, we found that the mRNA expression levels of lipogenic genes were not significantly changed after starvation. Conclusion: These findings suggest that in the initial stage of NEB in dairy cows, the liver coordinates an adaptive response by activating the IRE1 arm of the UPR to enhance ketogenesis, thereby avoiding a fatty liver status.

• 콘크리트학회논문집
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• 제21권3호
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• pp.291-302
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• 2009

현행 기준식에 따르면 초고강도콘크리트에서는 철근 인장이음길이보다 압축이음길이가 더 길어지는 현상이 발생된다. 초고강도콘크리트의 경제적 실용화를 위해 합리적인 압축이음강도의 평가가 필요하다. 이를 위해 압축이음의 거동 특성을 분석하고 영향인자를 도출하였으며, 설계강도 40, 60 MPa 콘크리트에 대한 압축이음 실험을 수행하였다. 압축이음강도는 부착과 지압으로 구성되고, 부착과 지압의 복합 거동에 의해 발현되므로, 압축이음 거동특성 및 강도평가를 위해서는 부착과 지압이 함께 존재하는 상태에서의 연구가 수행되어야한다. 인장이음과 달리 압축이음은 이음길이가 짧고 지압의 존재로 인해 콘크리트 강도의 영향이 크다. 실험결과 압축이음강도는 콘크리트의 제곱근에 비례하는 것으로 평가되었다. 부착과 지압 모두 주변 콘크리트의 응력상태에 따라 결정되는데, 콘크리트의 축방향 응력이 높기 때문에 철근 순간격 증가에 따른 이음강도 증가는 거의 없다. 지압강도는 이음길이와 철근 순간격에 무관하며, 콘크리트 강도의 제곱근의 함수로 표현할 수 있다. 파괴양상이 측면파열파괴와 유사하므로 지압강도는 앵커의 측면파열파괴 강도식을 활용하여 평가가 가능하다. 부착에 의해 발현되는 강도는 인장이음의 경우와 유사하므로, 인장이음강도에 비해 향상된 압축이음강도는 단부 지압효과로 설명될 수 있다.

• 한국강구조학회 논문집
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• 제22권5호
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• pp.477-485
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• 2010

다양한 모니터링 센서는 구조물의 손상측정과 예측에 많이 사용되고 있다. 광섬유센서, 압전소자(PZT) 센서, MEMS(Micro Electro Mechanical System)센서 등의 스마트 센서는 기존 센서를 대체하여 많은 분야에서 사용되고 있다. 본 논문에서는 PZT센서를 실험체에 부착한 후 충격하중을 가하여 PZT센서의 출력 전압의 특성을 이용하여 실험체의 손상을 예측하고자 하였다. PZT센서를 이용한 보 이음부의 손상실험에서는 $H-400{\times}200{\times}8{\times}13$ 철골을 이용하여 단순보를 제작하고 중앙에 볼트 이음을 하였으며, PZT센서의 민감도 측정을 위해 기존 가속도계의 가속도값과 PZT센서의 전압값의 FFT 결과를 비교하였다. 또한 이음부의 볼트풀림을 이용하여 단순보의 손상을 가정하고 손상계측실험도 병행하였다. 철골 플레이트 보의 손상계측 실험에서는 $PL600{\times}65{\times}5.8$로 단순보 실험체를 제작하여 세 곳에 손상을 주어 충격하중 실험을 하였다. 손상의 정도는 쇠톱을 이용하여 보 단면의 양쪽에 6~42 mm로 절단하였다. FFT를 사용하여 손상의 유무에 따른 고유진동수의 비(${\omega}_c/{\omega}$)를 구하여 손상을 파악하였고 모드에 따른 손상의 위치와 정도를 파악하였다.

• 한국가축번식학회지
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• 제20권4호
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• pp.371-378
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• 1997

본 연구는 인체 락토페린(hLF)을 우유 중으로 생산하는 형질전환 젖소의 개발에 관한 것이다. 이를 위한 모델 시스템으로서 락토페린 cDNAdhk 소의 베타-카제인 프로모터를 이용하여 형질전환 생쥐를 개발하였다. 발현 벡터의 락토페린에 대한 발현효율을 증가시키기 위하여 2개의 재조합 인트론을 삽입하였다. 20계통의 형질전환 생지를 개발하였는데 유즙에서의 락토페린 발현량은 1~200$\mu\textrm{g}$/ml이었다. hLF RNA의 발현 양상을 유선조직을 포함하여 뇌, 신장, 간 조직 등에서 조사하였을 때, 오직 유선에서만 발현되었을 뿐 아니라 엑손/인트론 경계 부위에서 정확하게 splicing되었다. hLF를 생산하는 형질전환 젖소를 개발하기 위하여 위에서 기술한 DNA를 소의 수정란에 미세주입한 후, 외과적 또는 비외과적 방법으로 대리모에 이식하였다. 한편, DNA가 주입된 수정란의 상태가 임신율에 미치는 영향을 조사하였다. 수정란을 최우수, 우수, 보통 등 3등급으로 나누었을 때, 각각의 임신율은 38.9, 15.4, 14.3%로 나타났다. 현재까지 유전자가 주입된 수정란을 대리모에 이식하여 태어난 35마리의 송아지 중, 30마리는 형절전환되지 않았으며 나머지는 현재 분석 중에 있다. 이상의 결과로 본 연구자들은 DNA가 미세주입된 젖소 수정란의 배양과 이식에 필요한 제반 기술을 확립하였으며, 아울러 임신율에 영향을 주는 여러 인자들에 대한 연구도 함께 조사하였다.