• Title/Summary/Keyword: vector analysis.

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Hepatitis B Virus-Induced TNF-a Expression in Hepa-lc1c7 Mouse Hepatoma Cell Line (마우스 Hepa-1c1c7 세포주에서 B형 간염 바이러스에 의한 tumor necrosis factor-a의 발현 유도)

  • Yea Sung Su;Jang Won Hee;Yang Young-Il;Lee Youn Jae;Kim Mi Seong;Seog Dae-Hyun;Park Yeong-Hong;Paik Kye-Hyung
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.38-44
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    • 2005
  • Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV, there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-1c1c7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-a, the inducibility of TNF-a expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-a expression. Infection of Hepa-1c1c7 cells with the serum of HBV carrier also increased TNF-a mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in $\beta-actin$ mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-lc1c7 in vitro and the viral infection induces TNF-a expression, which suggests that Hepa-lc1c7, a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.

Construction of Tomato yellow leaf curl virus Clones for Resistance Assessment in Tomato Plants (토마토 작물의 TYLCV 저항성 평가에 이용할 수 있는 감염성 클론 개발)

  • Choi, Seung Kook;Choi, Hak Soon;Yang, Eun Young;Cho, In Sook;Cho, Jeom Deog;Chung, Bong Nam
    • Horticultural Science & Technology
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    • v.31 no.2
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    • pp.246-254
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    • 2013
  • Five isolates of Tomato yellow leaf curl virus (TYLCV) collected from various regions of Korea were amplified using PCR and determined the sequences of full-length genome, respectively. The PCR-amplified DNA of each TYLCV isolate was introduced into a binary vector to construct infectious clone containing 1.9 copies of the corresponding viral genome. Various cultivars and breeding lines of tomato were inoculated with Agrobacterium tumefaciens harboring infectious clone of each TYLCV isolate to assess resistance against TYLCV. Susceptible cultivar 'Super-sunread' revealed typical yellowing and narrowing of the upper leaves. In contrast, breeding linesTY12, GC9, GC171, and GC173, which contained the TY-1 and/or TY-3 genes that confer resistance against TYLCV in nature, were completely symptomless, suggesting that the lines were resistant to challenging TYLCV isolates. Symptoms of TYLCV in susceptible tomato cultivars are significantly different from those of TYLCV in the resistant tomato cultivars at 30 days after agroinfiltration. Although genomic DNAs of TYLCV were detected from the breeding lines TY12, GC9, GC171, and GC173 using real-time PCR analysis with specific primers, levels of TYLCV DNA accumulation in the resistant breeding lines were much lower than those of TYLCV DNA accumulation in susceptible tomato cultivars. Similar symptom severity and levels of TYLCV DNA accumulation were observed from TYLCV infections mediated by Bemisia tabaci in the resistant and susceptible tomato cultivars. Concentration of agrobacterium did not affect the response of tomato cultivars against TYLCV inoculation. Taken together, these results suggest that TYLCV inoculation via agroinfiltration is as effective as inoculation through Bemisia tabaci and is useful for breeding programs of TYLCV-resistant tomato.

In Vitro Uptakes of Radiolabeled IVDU and IVFRU in Herpes Simplex Virus Type-1 Thymidine Kinase (HSV1-tk) Gene Transduced Morris Hepatoma Cell Line (단순 헤르페스 제 1형 티미딘 키나제 유전자 이입 간암세포주에서 방사표지 IVDU와 IVFRU의 섭취 평가)

  • Lee, Tae-Sup;Choi, Tae-Hyun;Ahn, Soon-Hyuk;Woo, Kwang-Sun;Jeong, Wee-Sup;Kwon, Hee-Chung;Awh, Ok-Doo;Choi, Chang-Woon;Lim, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.1
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    • pp.62-73
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    • 2004
  • Purpose: The herpes simplex virus type 1 thymidine kinase gene(HSV1-tk) is an attractive candidate as a reporter gene in noninvasive reporter gene monitoring system. The HSV1-tk gene was chosen as a reporter gene, because it has been extensively studied, and there are appropriate reporter probes, substrates of HSV1-tk gene product, to apply for HSV1-tk gene imaging. We used radiolabeled 5-iodovinyl-2'-deoxyuridine (IVDU) and 5-iodovinyl-2'-fluoro-2'-deoxyuridine (IVFRU) as reporter probes for HSV1-tk gene monitoring system. Materials and Methods: We prepared HSV1-tk gene transduced Morris hepatoma cell line using retroviral vector, MOLTEN containing HSV1-tk gene. And we confirmed the HSV1-tk gene expression by Northern blotting and Western blotting. We compared in vitro uptakes of radioiodinated IVDU and IVFRU to monitor HSV1-tk gene expression in Morris hepatoma cell line (MCA) and HSV1-tk gene tranduced MCA (MCA-tk) cells until 480 minutes. We also peformed correlation analysis between percentage of HSV1-tk gene tranduced MCA cell % (MCA-tk%) and uptakes of radiolabeled IVDU or IVFRU. Results: MCA-tk cell expressed HSV1-tk mRNA and HSV1-TK protein. Two compounds showed minimal uptake in MCA, but increased uptake was observed in MCA-tk. IVDU showed 4-fold higher accumulation than IVFRU at 480 min in MCA-tk (p<0.01). Both IVDU and IVFRU uptake were linearly correlated ($R^2>0.96$) with increasing MCA-tk%. Conclusion: The radiolabeld IVDU and IVFRU showed higher specific accumulation in retrovirally HSV1-tk gene transfected Morris hepatoma cell line. Both IVDU and IVFRU could be used as good substrates for evaluation of HSV1-tk gene expression.

Enhanced Growth Inhibition by Combined Gene Transfer of p53 and $p16^{INK4a}$ in Adenoviral Vectors to Lung Cancer Cell Lines (폐암세포주에 대한 p53 및 $p16^{INK4a}$의 복합종양억제유전자요법의 효과)

  • Choi, Seung -Ho;Park, Kyung-Ho;Seol, Ja-Young;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.67-75
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    • 2001
  • Background : Two tumor suppressor genes, p53 and p16, which have different roles in controlling the cell cycle and inducing apoptosis, are frequently inactivated during carcinogenesis including lung cancer. Single tumor suppressor gene therapies using either with p53 or p16 have been studied extensively. However, there is a paucity of reports regarding a combined gene therapy using these two genes. Methods : The combined effect of p53 and p16 gene transfer by the adenoviral vector on the growth of lung cancer cell lines and its interactive mechanism was investigated. Results : An isobologram showed that the co-transduction of p53 and p16 exhibited a synergistic growth in hibitory effect on NCI H358 and an additive effect on NCI H23. Cell cycle analysis demonstrated the induction of a synergistic G1/S arrest by a combined p53 and p16 transfer. This synergistic interaction was again confirmed in a soft agar confirmed in a soft agar clonogenic assay. Conclusion : These observations suggest the potential of a p53 and p16 combination gene therapy as another potent strategy in cancer gene therapy.

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The Determination Factor's Variation of Real Estate Price after Financial Crisis in Korea (2008년 금융위기 이후 부동산가격 결정요인 변화 분석)

  • Kim, Yong-Soon;Kwon, Chi-Hung;Lee, Kyung-Ae;Lee, Hyun-Rim
    • Land and Housing Review
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    • v.2 no.4
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    • pp.367-377
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    • 2011
  • This paper investigates the determination factors' variation of real estate price after sub-prime financial crisis, in korea, using a VAR model. The model includes land price, housing price, housing rent (Jensei) price, which time period is from 2000:1Q to 2011:2Q and uses interest rate, real GDP, consumer price index, KOSPI, the number of housing construction, the amount of land sales and practices to impulse response and variance decomposition analysis. Data cover two sub-periods and divided by 2008:3Q that occurred the sub-prime crisis; one is a period of 2000:1Q to 2008:3Q, the other is based a period of 2000:1Q to 2011:2Q. As a result, Comparing sub-prime crisis before and after, land price come out that the influence of real GDP is expanding, but current interest rate's variation is weaken due to the stagnation of current economic status and housing construction market. Housing price is few influenced to interest rate and real GDP, but it is influenced its own variation or Jensei price's variation. According to the Jensei price's rapidly increasing in nowadays, housing price might be increasing a rising possibility. Jensei price is also weaken the influence of all economic index, housing price, comparing before sub-prime financial crisis and it is influenced its own variation the same housing price. As you know, real estate price is weakened market basic value factors such as, interest rate, real GDP, because it is influenced exogenous economic factors such as population structural changes. Economic participators, economic officials, consumer, construction supplyers need to access an accurate observation about current real estate market and economic status.

Development of Marker-free Transgenic Rice for Increasing Bread-making Quality using Wheat High Molecular Weight Glutenin Subunits (HMW-GS) Gene (밀 고분자 글루테닌 유전자를 이용하여 빵 가공적성 증진을 위한 마커 프리 형질전환 벼의 개발)

  • Park, Soo-Kwon;Shin, DongJin;Hwang, Woon-Ha;Oh, Se-Yun;Cho, Jun-Hyun;Han, Sang-Ik;Nam, Min-Hee;Park, Dong-Soo
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1317-1324
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    • 2013
  • High-molecular weight glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the wheat grain. We have produced marker-free transgenic rice plants containing a wheat Glu-1Bx7 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using the Agrobacterium-mediated co-transformation method. The Glu-1Bx7-own promoter was inserted into a binary vector for seed-specific expression of the Glu-1Bx7 gene. Two expression cassettes comprised of separate DNA fragments containing only Glu-1Bx7 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to the Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Glu-1Bx7 or HPTII was infected to rice calli at a 3:1 ratio of Glu-1Bx7 and HPTII, respectively. Then, among 216 hygromycin-resistant $T_0$ plants, we obtained 24 transgenic lines with both Glu-1Bx7 and HPTII genes inserted into the rice genome. We reconfirmed integration of the Glu-1Bx7 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the wheat Glu-1Bx7 were stably expressed in the rice $T_1$ seeds. Finally, the marker-free plants harboring only the Glu-1Bx7 gene were successfully screened at the $T_1$ generation.

A Hierarchical Cluster Tree Based Fast Searching Algorithm for Raman Spectroscopic Identification (계층 클러스터 트리 기반 라만 스펙트럼 식별 고속 검색 알고리즘)

  • Kim, Sun-Keum;Ko, Dae-Young;Park, Jun-Kyu;Park, Aa-Ron;Baek, Sung-June
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.3
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    • pp.562-569
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    • 2019
  • Raman spectroscopy has been receiving increased attention as a standoff explosive detection technique. In addition, there is a growing need for a fast search method that can identify raman spectrum for measured chemical substances compared to known raman spectra in large database. By far the most simple and widely used method is to calculate and compare the Euclidean distance between the given spectrum and the spectra in a database. But it is non-trivial problem because of the inherent high dimensionality of the data. One of the most serious problems is the high computational complexity of searching for the closet spectra. To overcome this problem, we presented the MPS Sort with Sorted Variance+PDS method for the fast algorithm to search for the closet spectra in the last paper. the proposed algorithm uses two significant features of a vector, mean values and variance, to reject many unlikely spectra and save a great deal of computation time. In this paper, we present two new methods for the fast algorithm to search for the closet spectra. the PCA+PDS algorithm reduces the amount of computation by reducing the dimension of the data through PCA transformation with the same result as the distance calculation using the whole data. the Hierarchical Cluster Tree algorithm makes a binary hierarchical tree using PCA transformed spectra data. then it start searching from the clusters closest to the input spectrum and do not calculate many spectra that can not be candidates, which save a great deal of computation time. As the Experiment results, PCA+PDS shows about 60.06% performance improvement for the MPS Sort with Sorted Variance+PDS. also, Hierarchical Tree shows about 17.74% performance improvement for the PCA+PDS. The results obtained confirm the effectiveness of the proposed algorithm.

Automatic Speech Style Recognition Through Sentence Sequencing for Speaker Recognition in Bilateral Dialogue Situations (양자 간 대화 상황에서의 화자인식을 위한 문장 시퀀싱 방법을 통한 자동 말투 인식)

  • Kang, Garam;Kwon, Ohbyung
    • Journal of Intelligence and Information Systems
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    • v.27 no.2
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    • pp.17-32
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    • 2021
  • Speaker recognition is generally divided into speaker identification and speaker verification. Speaker recognition plays an important function in the automatic voice system, and the importance of speaker recognition technology is becoming more prominent as the recent development of portable devices, voice technology, and audio content fields continue to expand. Previous speaker recognition studies have been conducted with the goal of automatically determining who the speaker is based on voice files and improving accuracy. Speech is an important sociolinguistic subject, and it contains very useful information that reveals the speaker's attitude, conversation intention, and personality, and this can be an important clue to speaker recognition. The final ending used in the speaker's speech determines the type of sentence or has functions and information such as the speaker's intention, psychological attitude, or relationship to the listener. The use of the terminating ending has various probabilities depending on the characteristics of the speaker, so the type and distribution of the terminating ending of a specific unidentified speaker will be helpful in recognizing the speaker. However, there have been few studies that considered speech in the existing text-based speaker recognition, and if speech information is added to the speech signal-based speaker recognition technique, the accuracy of speaker recognition can be further improved. Hence, the purpose of this paper is to propose a novel method using speech style expressed as a sentence-final ending to improve the accuracy of Korean speaker recognition. To this end, a method called sentence sequencing that generates vector values by using the type and frequency of the sentence-final ending appearing in the utterance of a specific person is proposed. To evaluate the performance of the proposed method, learning and performance evaluation were conducted with a actual drama script. The method proposed in this study can be used as a means to improve the performance of Korean speech recognition service.

Study on the Safety of Firefly Luciferase in Human as a Transient Reporter Gene of Oncolytic Virotherapy (항암 바이러스 치료제의 보고유전자로써 반딧불이 루시퍼레이즈의 인체 내 안전성에 대한 연구)

  • Hong, Young Mi;Yoon, Woong Hee;Lee, You Ra;Kim, Soo Ji;Ngabire, Daniel;Narayanasamy, Badrinath;Ornella, Mefotse Saha Cyrelle;Kim, Myunghee;Cho, Euna;Lee, Bora;Hwang, Tae-Ho
    • Journal of Life Science
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    • v.31 no.11
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    • pp.1028-1036
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    • 2021
  • Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.

Application of spatiotemporal transformer model to improve prediction performance of particulate matter concentration (미세먼지 예측 성능 개선을 위한 시공간 트랜스포머 모델의 적용)

  • Kim, Youngkwang;Kim, Bokju;Ahn, SungMahn
    • Journal of Intelligence and Information Systems
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    • v.28 no.1
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    • pp.329-352
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    • 2022
  • It is reported that particulate matter(PM) penetrates the lungs and blood vessels and causes various heart diseases and respiratory diseases such as lung cancer. The subway is a means of transportation used by an average of 10 million people a day, and although it is important to create a clean and comfortable environment, the level of particulate matter pollution is shown to be high. It is because the subways run through an underground tunnel and the particulate matter trapped in the tunnel moves to the underground station due to the train wind. The Ministry of Environment and the Seoul Metropolitan Government are making various efforts to reduce PM concentration by establishing measures to improve air quality at underground stations. The smart air quality management system is a system that manages air quality in advance by collecting air quality data, analyzing and predicting the PM concentration. The prediction model of the PM concentration is an important component of this system. Various studies on time series data prediction are being conducted, but in relation to the PM prediction in subway stations, it is limited to statistical or recurrent neural network-based deep learning model researches. Therefore, in this study, we propose four transformer-based models including spatiotemporal transformers. As a result of performing PM concentration prediction experiments in the waiting rooms of subway stations in Seoul, it was confirmed that the performance of the transformer-based models was superior to that of the existing ARIMA, LSTM, and Seq2Seq models. Among the transformer-based models, the performance of the spatiotemporal transformers was the best. The smart air quality management system operated through data-based prediction becomes more effective and energy efficient as the accuracy of PM prediction improves. The results of this study are expected to contribute to the efficient operation of the smart air quality management system.