• Entomological Research
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• 제48권6호
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• pp.533-539
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• 2018

The RNA interference (RNAi) has been considered as an important genetic tool and applied to develop a new living modified (LM) crop trait which is an improvement of nutrient quality or pest management. The RNAi of DvSnf7 has been used for resistance to LM maize and the Western Corn Rootworm which is a major agricultural pest for the US Corn Belt. Most of the environmental risk assessments (ERA) of double strand RNA (dsRNA) have been performed using in vitro transcript products, and not in vivo expressed product. A large amount of dsRNA was required for the acute toxicity assay of water fleas. Therefore development of massive dsRNA purification techniques is critical. Daphnia, a freshwater microcrustacean, is a model organism for studying cellular and molecular mechanism involved in life history traits and ecotoxicology. In this study, we established the massive dsRNA purification method using Escherichia coli and implemented acute toxicity assays to Daphnia magna. As a result, the present RNase A and DNase I, dsRNA was efficiently purified without any special techniques or equipment. Even though purified dsRNA existed during the acute toxicity test, lethality or abnormal behavior were not observed in D. magna. These results indicated that GFP and DvSnf7 dsRNA were not significantly affected to D. magna due to their lack of sequence matching in its genome. The purification method of dsRNA and the acute toxicity assay of water fleas using purified dsRNA would be suitable for the toxicological studies of LMOs to aquatic non-target organisms.

• Advances in environmental research
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• 제7권3호
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• pp.213-223
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• 2018

Continuous input into the aquatic ecosystem and persistent structures have created concern of antibiotics, primarily due to the potential for the development of antimicrobial resistance. Degradation kinetics and mineralization of vancomycin B (VAN-B) by photocatalysis using $TiO_2$ and ZnO nanoparticles was monitored at natural pH conditions. Photocatalysis (PC) efficiency was followed by means of UV absorbance, total organic carbon (TOC), and HPLC results to better monitor degradation of VAN-B itself. Experiments were run for two initial VAN-B concentrations ($20-50mgL^{-1}$) and using two catalysts $TiO_2$ and ZnO at different concentrations (0.1 and $0.5gL^{-1}$) in a multi-lamp batch reactor system (200 mL water volume). Furthermore, a set of toxicity tests with Daphnia magna was performed to evaluate the potential toxicity of oxidation by-products of VAN-B. Formation of intermediates such as chlorides and nitrates were monitored. A rapid VAN-B degradation was observed in ZnO-PC system (85% to 70% at 10 min), while total mineralization was observed to be relatively slower than $TiO_2-PC$ system (59% to 73% at 90 min). Treatment efficiency and mechanism of degradation directly affected the rate of transformation and by-products formation that gave rise to toxicity in the treated samples.

• 식물병연구
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• 제25권1호
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• pp.1-7
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• 2019

벼도열병균은 벼를 재배하는 모든 지역에서 경제적으로 매우 중요한 병이다. 또한, 벼도열병균은 기주인 벼와 유전자 대유전자설이 적용되는 대표적인 식물병원균 모델이다. 재배지에 도입된 새로운 저항성 벼 품종의 빠른 저항성 상실은 병원균 집단의 레이스 변이가 주요 메커니즘으로 제안되고 있다. 이러한 새로운 레이스 변이는 저항성 유전자에 대항하는 비병원성 유전자의 변이에 의해 나타날 수 있는데, (i) 점돌연변이, (ii) 전이인자(transposon)의 삽입, (iii) frame shift등이 그 대표적인 예라고 할 수 있다. 비병원성 유전자 AVR-Pita1은 이러한 다양한 변이의 원인들이 모두 보고된 대표적인 비병원성 유전자이다. 이 총설에서는 비병원성 유전자 AVR-Pita1에 관한 다양한 정보를 제시하고, 상동성 유전자들인 AVR-Pita2 및 AVR-Pita3 유전자를 정리하였다. 이와 함께, 변이의 원인이 되는 다양한 예제를 리뷰 하였다.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.367-372
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• 2019

Deactivation of aminoglycosides by their modifying enzymes, including a number of aminoglycoside O-phosphotransferases, is the most ubiquitous resistance mechanism in aminoglycoside-resistant pathogens. Nonetheless, in a couple of biosynthetic pathways for gentamicins, fortimicins, and istamycins, phosphorylation of aminoglycosides seems to be a unique and initial step for the creation of a natural defensive structural feature such as a 3',4'-dideoxy scaffold. Our aim was to elucidate the biochemical details on the beginning of these C3',4'-dideoxygenation biosynthetic steps for aminoglycosides. The biosynthesis of istamycins must surely involve these 3',4'-didehydroxylation steps, but much less has been reported in terms of characterization of istamycin biosynthetic genes, especially about the phosphotransferase-encoding gene. In the disruption and complementation experiments pointing to a putative gene, istP, in the genome of wild-type Streptomyces tenjimariensis, the function of the istP gene was proved here to be a phosphotransferase. Next, an in-frame deletion of a known phosphotransferase-encoding gene forP from the genome of wild-type Micromonospora olivasterospora resulted in the appearance of a hitherto unidentified fortimicin shunt product, namely 3-O-methyl-FOR-KK1, whereas complementation of forP restored the natural fortimicin metabolite profiles. The bilateral complementation of an istP gene (or forP) in the ${\Delta}forP$ mutant (or ${\Delta}istP$ mutant strain) successfully restored the biosynthesis of 3',4'-dideoxy fortimicins and istamycins, thus clearly indicating that they are interchangeable launchers of the biosynthesis of 3',4'-dideoxy types of 1,4-diaminocyclitol antibiotics.

• 정보보호학회논문지
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• 제31권4호
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• pp.661-674
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• 2021

커널을 변조하는 루트킷과 같은 악성코드가 만약 메모리 분석을 회피하기 위한 메커니즘을 추가하게 될 경우, 분석이 어려워지거나 불가능하게 되면서 분석가의 판단에 악영향을 미칠 수 있다. 따라서 향후 고도화된 커널 변조를 통해 탐지를 우회하는 루트킷과 같은 악성코드에 선제적으로 대응하고자 한다. 이를 위해 공격자의 관점에서 윈도우 커널에서 사용되는 주요 구조체를 분석하고, 커널 객체를 변조할 수 있는 방법을 적용하여 메모리 덤프 파일에 변조를 진행하였다. 변조 결과 널리 사용되는 메모리 분석 도구에서 탐지가 되지 않는 것을 실험을 통해 확인하였다. 이후 분석가의 관점에서 변조 저항성의 개념을 사용하여 변조를 탐지할 수 있는 소프트웨어 형태로 만들어 기존 메모리 분석 도구에서 탐지되지 않는 영역에 대해 탐지 가능함을 보인다. 본 연구를 통해 선제적으로 커널 영역에 대해 변조를 시도하고 정밀 분석이 가능하도록 인사이트를 도출하였다는 데 의의가 있다 판단된다. 하지만 정밀 분석을 위한 소프트웨어 구현에 있어 필요한 탐지 규칙을 수동으로 생성해야 한다는 한계점이 존재한다.

• Journal of Ginseng Research
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• 제45권1호
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• pp.86-97
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• 2021

Background: Panax ginseng Meyer has been used as a nourishing edible herb in East Asia for thousands of years. 25-OH-PPT was first discovered as a natural rare triterpenoid saponin in ginseng stems and leaves by our group. Research found that it showed strong inhibitory effects on α-glucosidase and protein tyrosine phosphatase 1B, and protected cardiocytes (H9c2) through PI3K/Akt pathway. Methods: In the research, in order to optimize the 25-OH-PPT enrichment process, optimal macroporous resins and optimal purification conditions were studied. Meanwhile, the hypoglycemic effect and mechanism of 25-OH-PPT were evaluated by using STZ to establish insulin-dependent diabetic mice and the spontaneous type 2 diabetes DB/DB mice. Results and Conclusion: Research found that 25-OH-PPT can reduce blood glucose and enhance glucose tolerance in STZ model mice. It increases insulin sensitivity by upregulating GLUT4 and AMPK in skeletal muscle, and activating insulin signaling pathways. In DB/DB mice, 25-OH-PPT achieves hypoglycemic effects mainly by activating the insulin signaling pathway. Meanwhile, through the influence of liver inflammatory factors and lipids in serum, it can be seen that 25-OH-PPT has obvious anti-inflammatory and lipid-lowering effects. These results provide new insights into the study of ginseng as a functional food.

• 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.

• Biomolecules & Therapeutics
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• 제30권1호
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• pp.19-27
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• 2022

Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase widely expressed in many cancers such as non-small cell lung cancer (NSCLC), pancreatic cancer, breast cancer, and head and neck cancer. Mutations such as L858R in exon 21, exon 19 truncation (Del19), exon 20 insertions, and others are responsible for aberrant activation of EGFR in NSCLC. First-generation EGFR tyrosine kinase inhibitors (TKIs) such as gefitinib and erlotinib have clinical benefits for EGFR-sensitive (L858R and Del19) NSCLC patients. However, after 10-12 months of treatment with these inhibitors, a secondary T790M mutation at the gatekeeper position in the kinase domain of EGFR was identified, which limited the clinical benefits. Second-generation EGFR irreversible inhibitors (afatinib and dacomitinib) were developed to overcome this T790M mutation. However, their lack of selectivity toward wild-type EGFR compromised their clinical benefits due to serious adverse events. Recently developed third-generation irreversible EGFR TKIs (osimertinib and lazertinib) are selective toward driving mutations and the T790M mutation, while sparing wild-type EGFR activity. The latest studies have concluded that their efficacy was also compromised by additional acquired mutations, including C797S, the key residue cysteine that forms covalent bonds with irreversible inhibitors. Because second- and third-generation EGFR TKIs are irreversible inhibitors, they are not effective against C797S containing EGFR triple mutations (Del19/T790M/C797S and L858R/T790M/C797S). Therefore, there is an urgent unmet medical need to develop next-generation EGFR TKIs that selectively inhibit EGFR triple mutations via a non-irreversible mechanism.

• BMB Reports
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• 제54권12호
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• pp.608-613
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• 2021

Melanoma, the most serious type of skin cancer, exhibits a high risk of metastasis. Although chemotherapeutic treatment for metastatic melanoma improves disease outcome and patient survival, some patients exhibit resistance or toxicity to the drug treatment regime. OTUB1 is a deubiquitinating enzyme overexpressed in several cancers. In this study, we investigated the effects of inhibiting OTUB1 expression on melanoma-cell proliferation and viability and identified the underlying molecular mechanism of action of OTUB1. We did endogenous OTUB1 knockdown in melanoma cells using short interfering RNA, and assessed the resulting phenotypes via MTT assays, Western blotting, and cell-cycle analysis. We identified differentially expressed genes between OTUB1-knockdown cells and control cells using RNA sequencing and confirmed them via Western blotting and reverse transcription polymerase chain reaction. Furthermore, we investigated the involvement of apoptotic and cell survival signaling pathways upon OTUB1 depletion. OTUB1 depletion in melanoma cells decreased cell viability and caused simultaneous accumulation of cells in the sub-G1 phase, indicating an increase in the apoptotic-cell population. RNA sequencing of OTUB1-knockdown cells revealed an increase in the levels of the apoptosis-inducing protein TRAIL. Additionally, OTUB1-knockdown cells exhibited increased sensitivity to PLX4032, a BRAF inhibitor, implying that OTUB1 and BRAF act collectively in regulating apoptosis. Taken together, our findings show that OTUB1 induces apoptosis of melanoma cells in vitro, likely by upregulating TRAIL, and suggest that approaches targeting OTUB1 can be developed to provide novel therapeutic strategies for treating melanoma.

• Journal of the Korean Wood Science and Technology
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• 제32권2호
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• pp.50-57
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• 2004

본 실험은 건조되지 않은 목재를 고온의 아마인유에 침지처리함으로써 목재건조와 동시에 발수효과를 얻고자 실시하였다.함수율 90% 이상의 소나무생재를 온도 150℃의 아마인유에서 6시간 침지처리하였을 때 목재함수율은 10%까지 떨어졌으며, 목재 축방향의 깊이에 관계없이 목재단면의 20%에 해당하는 아마인유가 침투하였다. 이때 목재의 강도적 손실은 발생하지 않는 것으로 나타났다. 목재표면의 천공처리는 목재의 내부할열방지에 효과적이었고, 아마인유의 깊은 침투를 유도할 수 있었다. 흡수성 시험 결과 처리재는 무처리재에 비하여 높은 발수효과를 나타냈다. 변색균 및 부후균에 대한 항균효력시험은 처리재 자체로서 균에 대한 독성 기작이 없었으나, 백색부후균 및 갈색부후균에 대해서는 무처리재보다 낮은 중량감소율을 나타내는 등 방부효력의 상승이 있었다.