• The Plant Pathology Journal
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• 제22권2호
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• pp.139-146
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• 2006

The 3' region of Potato virus X (PVX) has the 74 nt 3'-nontranslated region (NTR) that is conserved among all potexviruses and contains several cis-acting elements for minus-strand and plus-strand RNA accumulation. Three stem-loop structures (SL1-SL3), especially formation of SL3 and U-rich sequence of SL2, and near upstream elements in the 3' NTR were previously demonstrated as important cis-acting elements. To Investigate the binding of these cis-acting elements within 3' end with host protein, we used the electrophoretic mobility shift assays (EMSA) and UV-cross linking analysis. The EMSA with cellular extracts from tobacco and RNA transcripts corresponding to the 150 nt of the 3' end of PVX RNA showed that the 3' end of PVX formed complexes with cellular proteins. The specificity of protein binding was confirmed through competition assay by using with 50-fold excess of specific and non-specific probes. We also conducted EMSA with RNAs containing various mutants on those cis-acting elements (${\Delta}10$10, SL3B, SL2A and ${\Delta}21$; J Mol Biol 326, 701-720) required for efficient PVX RNA accumulation. These analyses supported that these cis-acting elements are required for interaction with host protein(s). UV-cross linking analysis revealed that at least three major host proteins of about 28, 32, and 42 kDa in mass bound to these cis-elements. These results indicate that cis-acting elements from 3' end which are important for minus and plus-strand RNA accumulation are also required for host protein binding.

• 한국잠사곤충학회지
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• 제35권2호
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• pp.129-133
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• 1993

Bacillus thruingiensis var. kurstaki HD-1의 내독소 단백질 유전자의 발현기작을 규명하기 위하여 이 균으로부터 내독소 단백질 유전자가 존재하는 것으로 확인된 29Md와 44Md plasmid를 분리한 후, Sau3AI 제어효소로 부분절단하고, pBR322 BamHI site에 ligation하여, E. coli HB101 strain에 transformation시켜, 3,000여개의 Ampr/Ters한 colony를 얻어 면역학적 방법과 살충적 검정으로 통해 재조합 균주 KC1을 얻었다. PKC1 plasmid DNA는 vector DNA를 포함하여 약 12kb 정도의 크기를 가지며, B. t k HD-1 내독소 단백질과 이동도가 같거나 일치하는 132kd, 117kd의 KC1 specific band 2개를 얻었다. KC1 cell extract를 첨식한 결과 약 80% 치사율을 보였다.

• 한국환경과학회지
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• 제19권3호
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• pp.379-387
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• 2010

Sediments of Little Scioto (LS) River in Ohio was contaminated by poor disposal of creosote from Baker Wood Creosoting Facility. Among the primary compounds of creosote, Polycyclic Aromatic Hydrocarbons (PAHs) are the most common ingredient PAHs are known for toxic, carcinogenic and mutagenic compounds. There are many difficulties to remove the PAHs in nature environment because their characteristics are having a less water-solubility, volatile and low mobility properties as increasing the molecular weight. The generation of hydroxyl radicals (${\cdot}OH$) and hydrogen peroxide ($H_2O_2$) forms as well as high temperature (5000 K) and pressure (1000 atm) by a physico-chemical effects of ultrasound during a cavitation collapse can promote the degradation and desorption of PAHs in sediment And it can also produces shock wave and microjets which are able to change the size and surface of particle in solid-liquid system as one of physical effects. Therefore, we explored to understand the role of particle size, the effect of elimination for PAHs concentration by ultrasound and optimize the conditions for ultrasonic treatment. The condition of various size of particles (> $150{\mu}m$, < $150{\mu}m$) and solid-liquid ratio (12.5g/L, 25g/L) for the treatment was considered and ultrasonic power (430 W/L) with liquid - hexane extraction and microwave extraction method were applied after ultrasound treatment.

• Asian Pacific Journal of Cancer Prevention
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• 제16권12호
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• pp.4981-4985
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• 2015

Background: Colorectal cancer (CRC) is a leading cause of morbidity and mortality worldwide. Targeting autophagic cell death is emerging as a novel strategy in cancer chemotherapy. Aldose reductase (AR) catalyzes the rate limiting step of the polyol pathway of glucose metabolism; besides reducing glucose to sorbitol, AR reduces lipid peroxidation-derived aldehydes and their glutathione conjugates. A complex interplay between autophagic cell death and/or survival may in turn govern tumor metastasis. This exploratory study aimed to investigate the potential role of AR inhibition using a novel inhibitor Fidarestat in the regulation of autophagy in CRC cells. Materials and Methods: For glucose depletion (GD), HT-29 and SW480 CRC cells were rinsed with glucose-free RPMI-1640, followed by incubation in GD medium +/- Fidarestat ($10{\mu}M$). Proteins were extracted by a RIPA-method followed by Western blotting ($35-50{\mu}g$ of protein; n=3). Results: Autophagic regulatory markers, primarily, microtubule associated protein light chain (LC) 3, autophagy-related gene (ATG) 5, ATG 7 and Beclin-1 were expressed in CRC cells; glyceraldehyde-3 phosphate dehydrogenase (GAPDH) was used as an internal reference. LC3 II (14 kDa) expression was relatively high compared to LC3A/B I levels in both CRC cell lines, suggesting occurrence of autophagy. Expression of non-autophagic markers, high mobility group box (HMG)-1 and Bcl-2, was comparatively low. Conclusions: GD +/- ARI induced autophagy in HT-29 and SW-480 cells, thereby implicating Fidarestat as a promising therapeutic agent for colorectal cancer; future studies with more potent ARIs are warranted to fully dissect the molecular regulatory networks for autophagy in colorectal carcinoma.

• BMB Reports
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• 제35권3호
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• pp.273-282
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• 2002

Sp3 is a bifunctional transcription factor that has been reported to stimulate or repress the transcription of numerous genes. Although the size of Sp3 mRNA is 4.0kb, the size of the known Sp3 cDNA sequence is 3.6kb. Thus, Sp3 functional studies have been performed with an artificially introduced start codon, and thus an amino-terminus that differs from the wild-type. Ideally, full-length cDNA expression vectors with the appropriate start codon should be utilized for these studies. Using 5'rapid amplification of cDNA ends, a full-length Sp3 cDNA clone was generated and the sequence verified in nine cell lines. No AUG initiation codon was present. However, stop codons were present in all three frames 5' to the known coding sequence. In vitro translation of this full-length cDNA clone produced the expected three isoforms-one at 100 kDa and two in the mid 60 kDa range. Electrophoretic mobility shift assays showed that the protein products had the ability to bind to the Sp1/3 consensus sequence. In vitro studies, using our Sp3 clone and site directed mutagenesis, identified the translation initiation site for the larger isoform as AUA. AUA has not been previously described as an endogenous initiation codon in eukaryotes.

• BMB Reports
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• 제31권1호
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• pp.64-69
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• 1998

Recent studies have suggested that integrin (CR3) participates in the signal transduction pathways of certain GPI-anchored phagocytic receptors including $Fc{\gamma}RIIIB$. One consequence of this functional linkage is an inducible association between CR3 and cortical microfilaments that is triggered by $Fc{\gamma}RIIIB$ binding to immobilized immune complexes (IC). That this signaling event requires the co-expression of $Fc{\gamma}RIIIB$ with CR3 was documented by the use of NIH 3T3 transfectants expressing both CR3 and $Fc{\gamma}RIIIB$ (clone 3-23), CR3 alone (clone 3-19), and $Fc{\gamma}RIIIB$ alone (clone 3-15). Pretreatment of 3-23 cells with protein kinase inhibitors such as staurosporine and methyl 2,5-dihydroxycinnamate (MDHC) blocked IC-stimulated CR3 microfilament proximity without affecting the extent to which $Fc{\gamma}RIIIB$ constrains the lateral membrane mobility of a subset of CR3 on the cell surface (as measured in fluorescence recovery after photobleaching experiments). These data support that CR3 and $Fc{\gamma}RIIIB$ molecules are physically and functionally associated and that ligation of FcgRIIIB triggers CR3-dependent signal transduction.

• 대한한의학회지
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• 제24권2호
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• pp.179-192
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• 2003

Objectives : Boyanghwanoh-tang (Buyanhaiwu-tang) has been used as a prescription for stroke, senile and vascular dementia, ischemic brain and heart damage in Oriental traditional medicine. However, there is little known about the mechanism by which the water extracts of Boyanghwanoh-tang (Buyanhaiwu-tang) rescue cells fromthese damages, and little is known about the protective mechanisms of Boyanghwanoh-tang (Buyanhaiwu-tang) on oxidative stress in neuronal cells. Therefore, we have investigated the role of Boyanghwanoh-tang (Buyanhaiwu-tang) on serum and glucose deprived apoptosis in PC12 cells. Methods : PC12 Cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MIT assay. We used DNA fragmentation and caspase 1, 2, 3, 6, 9-likeproteases activation assay. Transcriptional activation of NF-kB was assessed by using electrophoretic mobility shift assay. Results : Boyanghwanoh-tang (Buyanhaiwu-tang) rescued PC12 cells from apoptotic death by serum and glucose deprivation in a dose-dependent manner. The nuclear staining of PC12 cells clearly showed that Boyanghwanoh-tang (Buyanhaiwu-tang) attenuated nuclear condensation and fragmentation, which represent typical neuronal apoptotic characteristics. Boyanghwanoh-tang (Buyanhaiwu-tang) also prevents fragmentation of genomic DNA and activation of caspase 3-like protease in serum and glucose deprived PC12 cells. Furthermore, Boyanghwanoh-tang (Buyanhaiwu-tang) reduced the activation of NF-kB by serum and glucose-deprived apoptosis. Conclusions : These findings suggest that serum and glucose deprivation induces reduced glutathione (GSH) depletion, and consequently, apoptosis through endogenously produced reactive oxygen species in PC12 cells. Also, our data indicated that Boyanghwanoh-tang (Buyanhaiwu-tang) has protective effects against the serum and glucose deprived deaths of PC12 cells, which are mediated by the generation of GSH that, in turn, can reduce oxidative stress caused by reactive oxygen species (ROS) such as hydrogen peroxide.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5533-5537
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• 2013

MicroRNA-10b (miR-10b) has been reported to play an important role in some types of cancer, but the effects and possible mechanisms of action of miR-10b in the metastasis of nasopharyngeal carcinoma cells (NPC) have not been explored. The aim of the present study was to investigate the function of miR-10b in nasopharyngeal carcinoma and to determine the molecular mechanisms underlying its action. The MTT assay was used to assess proliferation of CNE-2Z cells. Wound healing and transwell migration assays were applied to assess cell migration and invasion, while and expression of E-cadherin and MMP-9 were detected using Western blot analysis. Real-time PCR was employed to detect the expression of genes related to migration and invasion and the $2^{-{\Delta}{\Delta}Ct}$ method was used to calculate the degree of expression. MTT assay showed the expression of miR-10b to have no effect on the proliferation of NPC cell lines. The wound healing assay showed that miR-10b mimics promoted the mobility and invasion of NPC cell lines. Inhibitors of miR-10b reduced the ability of NPC cell lines to migrate and invade. In addition, the expression of genes related to migration and invasion, such as E-cadherin, vimentin, and MMP-9, were confirmed to be different in the CNE-2Z NPC cell line transfected with miR-10b mimics and with miR-10b inhibitors. In the present study, miR-10b was found to upregulate the expression of MMP-9 and knockdown of miR-10b was found to significantly downregulate the expression of E-cadherin. On the whole, these results showed that miR-10b plays an important role in the invasion and metastasis of NPC cells.

• Natural Product Sciences
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• 제17권3호
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• pp.250-255
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• 2011

The immune system is finely balanced by the activities of pro-inflammatory and anti-inflammatory mediators or cytokines. Unregulated activities of these mediators can lead to the development of various inflammatory diseases. A variety of safe and effective anti-inflammatory agents are available with many more drugs under development. Of the natural compounds, the sesquiterpenes (nootkatone, ${\alpha}$-cyperone, valencene and ${\alpha}$-selinene) isolated from C. rotundus L. have received much attention because of their potential antiinflammatory effects. However, limited studies have been reported regarding the influence of sesquiterpene structure on anti-inflammatory activity. In the present study, the anti-inflammatory potential of four structurally divergent sesquiterpenes was evaluated in lipopolysaccaride (LPS)-stimulated RAW 264.7 cells, murine macrophages. Among the four sesquiterpenes, ${\alpha}$-cyperone and nootkatone, showed stronger anti-inflammatory and a potent NF-${\kappa}B$ inhibitory effect on LPS-stimulated RAW 264.7 cells. Molecular analysis revealed that various inflammatory enzymes (iNOS and COX-2) were reduced significantly and this correlated with downregulation of the NF-${\kappa}B$ signaling pathway. Additionally, electrophoretic mobility shift assays (EMSA) elucidated that nootkatone and ${\alpha}$-cyperone dramatically suppressed LPS-induced NF-${\kappa}B$-DNA binding activity using 32Plabeled NF-${\kappa}B$ probe. Hence, our data suggest that ${\alpha}$-cyperone and nootkatone are potential therapeutic agents for inflammatory diseases.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2011년도 제40회 동계학술대회 초록집
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• pp.152-152
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• 2011

Silicon 기반의 환경에서 연구 및 제조되는 전자소자는 반도체의 기술이 발전함에 따라 chip 선폭의 크기가 30 nm에서 20 nm, 그리고 그 이하의 크기로 점점 더 작아지는 요구에 직면하고 있다. 탄소나노 구조와 나노와이어 기술이 Silicon을 대신할 다음세대 기술로 주목받고 있다. 많은 연구결과들 중에서 III-V CMOS가 가장 빠른 접근 방법이라 예상한다. III-V족 물질을 이용하면 electron 보다 수십 배 이상의 이동도를 얻을 수 있으나 p-type의 구조를 구현하는 것이 해결해야 할 문제이다. p-type 3-5 족 화합물을 이용하여 에너지 밴드 갭의 변화를 가능하게 한다면 hole의 이동도를 크게 향상시킬 수 있어 silicon 기반의 p-type 소자보다 2~3배 더 빠른 소자의 구현이 가능하다. 3-5족 화합물 반도체의 성장 기술이 많이 진보되어 이를 이용하여 고속 소자를 구현한다면 시기적으로 더욱 빨리 다가올 것이라 예측한다. 에너지 밴드갭의 변화와 격자 부정합을 고려하여 SI InP 기판에 GaSb 물질을 채널로 사용한 p-type 2-dimensional hole gas (2DHG) 소자를 구현하였다. 관찰된 소자 구조의 박막 상태의 특징을 보이며 10 um ${\times}$ 10 um AFM 측정결과 1 nm 이하의 표면 거칠기를 가지며 상온에서의 hole 이동도는 약 650 cm2/Vs이고 sheet carrier density는 $5{\times}1012$ /cm2의 결과를 확인하였다. 실험결과 InP 기판위에 채널로 사용된 GaSb 박막을 올리는데 있어 가장 중요한 것은 Phosphorus, Arsenic, 그리고 Antimony 물질의 양과 이들의 변화시간의 조절이다. 본 발표에서 Semi-insulating InP 기판위에 electron이 아닌 hole을 반송자로 이용한 차세대 고속 전자소자를 구현하고자 하여 MBE (Molecular Beam Epitaxy)로 p-type 소자를 구현하여 실험하였다. 아울러 더욱 빠른 소자의 구현을 위하여 세계의 유수 그룹들의 연구 결과들과 앞으로 예상되는 고속 소자에 대해서 비교와 함께 많은 기술에 대해 논의하고자 한다.