• Journal of Microbiology and Biotechnology
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• 제19권9호
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• pp.1070-1076
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• 2009

Telomerase reverse transcriptase (TERT), which prolongs the replicative life span of cells, is highly upregulated in 85-90% of human cancers, whereas most normal somatic tissues in humans express limited levels of the telomerase activity. Therefore, TERT has been a potential target for anticancer therapy. Recently, we described a new approach to human cancer gene therapy, which is based on the group I intron of Tetrahymena thermophila. This ribozyme can specifically mediate RNA replacement of human TERT (hTERT) transcript with a new transcript harboring anticancer activity through a trans-splicing reaction, resulting in selective regression of hTERT-positive cancer cells. However, to validate the therapeutic potential of the ribozyme in animal models, ribozymes targeting inherent transcripts of the animal should be developed. In this study, we developed a Tetrahymena-based trans-splicing ribozyme that can specifically target and replace the mouse TERT (mTERT) RNA. This ribozyme can trigger transgene activity not only also in mTERT-expressing cells but hTERT-positive cancer cells. Importantly, the ribozyme could selectively induce activity of the suicide gene, a herpes simplex virus thymidine kinase gene, in cancer cells expressing the TERT RNA and thereby specifically hamper the survival of these cells when treated with ganciclovir. The mTERT-targeting ribozyme will be useful for evaluation of the RNA replacement approach as a cancer gene therapeutic tool in the mouse model with syngeneic tumors.

• 식물병연구
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• 제26권4호
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• pp.202-209
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• 2020

유기농업은 지속가능한 농업을 위해서 필요하며, 생물 다양성 유지를 위해 필수적인 경작 방법으로 전 세계적으로 성장하고 있는 농업 분야 중 하나이다. 유기농업은 인류의 건강과 환경에 유해한 영향을 끼질 수 있는 화학적 제재의 사용을 줄이고, 친환경적 동·식물성 비료, 식물 추출물, 미생물 제재 등이 사용되기 때문에 소비자에게 안전한 농산물을 제공할 수 있다. 노지 고추를 대상으로 3년간 유기농업 환경과 관행농업 환경에서 주요 병해 발생과 토양 미생물상의 조사하였다. 토양 미생물상의 차이는 차세대 염기서열 분석 기법을 활용하여 조사하였으며, 유기농업에 사용되는 제제의 종류에 따라 토양 미생물상이 다르게 형성되었다. 바이러스, 탄저병에 대해서는 관행농업에서의 병 발생률이 낮았으나, 청고병은 미생물 제재 사용에 의해 병 발생률이 낮게 나타났다. 미생물 제재를 사용한 토양에서 투입된 미생물이 토양 미생물에서 검출되었으며, 이는 미생물이 실제 토양에 정착했음을 시사한다.

• Nutrition Research and Practice
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• 제10권1호
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• pp.3-10
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• 2016

BACKGROUND/OBJECTIVES: Oligonol, mainly found in lychee fruit, is an antioxidant polyphenolic compound which has been shown to have anti-inflammatory and anti-cancer properties. The detailed mechanisms by which oligonol may act as an anti-aging molecule have not been determined. MATERIALS/METHODS: In this study, we evaluated the ability of oligonol to modulate sirtuin (SIRT) expression in human lung epithelial (A549) cells. Oligonol was added to A549 cells and reactive oxygen species production, mitochondrial superoxide formation, and p21 protein levels were measured. Signaling pathways activated upon oligonol treatment were also determined by western blotting. Furthermore, the anti-aging effect of oligonol was evaluated ex vivo in mouse splenocytes and in vivo in Caenorhabditis elegans. RESULTS: Oligonol specifically induced the expression of SIRT1, whose activity is linked to gene expression, metabolic control, and healthy aging. In response to influenza virus infection of A549 cells, oligonol treatment significantly up-regulated SIRT1 expression and down-regulated viral hemagglutinin expression. Oligonol treatment also resulted in the activation of autophagy pathways and the phosphorylation of AMP-activated protein kinase (AMPK). Furthermore, oligonol-treated spleen lymphocytes from old mice showed increased cell proliferation, and mRNA levels of SIRT1 in the lungs of old mice were significantly lower than those in the lungs of young mice. Additionally, in vivo lethality assay revealed that oligonol extended the lifespan of C. elegans infected with lethal Vibrio cholerae. CONCLUSIONS: These data demonstrated that oligonol may act as an anti-aging molecule by modulating SIRT1/autophagy/AMPK pathways.

• Journal of Animal Science and Technology
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• 제63권4호
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• pp.864-871
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• 2021

Infectious calf diarrhea is one of the most significant diseases of neonatal calves. This study is conducted to identify the prevalence of pathogens in calf diarrhea for 2 years. A total of 544 feces samples from Korean native beef calves were obtained to investigate selected seven pathogens causing calf diarrhea: bovine rotavirus, bovine coronavirus, Cryptosporidium parvum, bovine viral diarrhea virus, Eimeria species, Escherichia coli K99, and Salmonella species. The presence of diarrhea, the number and species of detected pathogens, and the calves' ages were analyzed using various statistical methods depending on the case. Of the 544 calves, 340 calves (62.5%) had normal feces and 204 calves (37.5%) had diarrhea. The presence of pathogens was significantly associated with diarrhea (p < 0.01) and fecal scores and the number of detected pathogens showed a significant linear trend (p < 0.001). Of the 7 target pathogens, 6 were detected in samples, but only C. parvum (p = 0.001) and bovine rotavirus (p < 0.001) were found at significantly higher rates in diarrheic calves than in non-diarrheic calves. Only Eimeria spp. showed a significant linear trend between the detection rate of the pathogen and the age groups (p < 0.05).

• Journal of Plant Biotechnology
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• 제47권2호
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• pp.157-163
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• 2020

급속한 산업화와 인구증가에 따른 심각한 환경과 식량문제는 인류의 생존을 위협하는 가장 중요한 현안문제로 대두되고 있다. 또한, 인구 증가에 따른 식량부족을 해결하기 위하여 농약과 화학비료의 무분별한 사용으로 인하여 농토는 산성화되어 황폐화가 되고 있고 먹이사슬 및 자연생태계의 파괴는 더 많은 농약의 사용을 필요하고 있다. 과다한 농약 사용으로 경제적인 부담의 가중과 잔류 농약으로 인한 소비자의 건강을 위협하고 있다. 또한, 증가된 화석연료의 사용은 공기중의 이산화탄소를 증가시켜 지구 온도의 상승을 초래하고 있으며, 결과적으로 기상이변, 지구온난화 및 사막화 등의 심각한 환경문제를 초래했다. 특히, 서늘한 기온에서 잘 자라는 배추, 무우, 감자 등의 고령지 농작물의 질적인 저하를 초래하여 피해가 증가하고 있지만, 최근들어 감자와 같은 알카리성 건강식품의 붐으로 수요가 증가되고 있다. 본 연구에서 환경스트레스에 관여하는 대두의 특이적인 칼모듈린, GmCaM-4 유전자를 감자에 과발현 시켜서 PR 유전자들의 발현이 지속적으로 유지되어 식물방어 기작이 활성화 되었음을 확인하였다. 또한, 그 형질전환 식물체는 초민감성 세포사멸 현상을 보였고, 무름병을 일으키는 병원균인 Erwinia carotovora subsp. Carotovora (Ecc)를 이용하여 GmCaM-4가 과발현된 감자에서 병 저항성이 증가하는 것을 확인 하였다. 최종적으로 지금까지 많이 연구되고 보고된 유전자원인 대두의 GmCaM-4 유전자를 활용하여 주요 식량자원인 감자에서 과발현 형질전환 식물체를 확보하여 다양한 병 저항성 증가를 통한 작물 생산성 향상에 매우 우수한 기술로 기대되는 바이다.

• Journal of Plant Biotechnology
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• 제38권1호
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• pp.62-68
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• 2011

원예작물의 생육을 저하시키는 각종 병충해로 인한 과도한 농약과 화학비료의 사용은 환경오염뿐만 아니라 작물의 생산량에도 큰 영향을 미치고 있다. 식물생명공학기술을 이용하여 농약이나 화학비료 사용량을 획기적으로 줄일 수 있는 식물체의 개발, 즉 형질전환을 이용한 분자 육종기술은 병충해 내성 농작물을 개발하여 과도한 화학비료의 사용에 따르는 여러가지 문제점들을 극복할 수 있는 대안으로 대두되고 있다. 본 연구에서는 모델식물인 애기장대에서 분리한 식물생체방어 신호전달에 관련된 AtCBP63 유전자를 감자에 과발현시켰고, 이러한 형질 전환 감자에서 병저항성에 관여하는 유전자인 PR-2, PR-3, PR-5 유전자들의 발현이 증가되어 지속적으로 식물 방어 기작이 활성화되어 있음을 확인하였다. 또한, 감자에서 무름병 (soft rot disease)을 일으켜 막대한 피해를 유발하는 병원성 세균인 Erwinia carotovora subsp. Carotovora (Ecc)를 이용하여 AtCBP63 유전자를 과발현한 감자에 감염시켰을 때, 병 저항성이 증가한다는 사실을 검증하였다. 앞으로, 다양한 곰팡이 균에 대응하여 AtCBP63 유전자를 과발현한 감자에 저항성을 검증하고자 한다.

• Journal of Veterinary Science
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• 제24권4호
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• pp.51.1-51.17
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• 2023

Background: To date, various genotypes of infectious bronchitis virus (IBV) have co-circulated and in Korea, GI-15 and GI-19 lineages were prevailing. The spike protein, particularly S1 subunit, is responsible for receptor binding, contains hypervariable regions and is also responsible for the emerging of novel variants. Objective: This study aims to investigate the putative major amino acid substitutions for the variants in GI-19. Methods: The S1 sequence data of IBV isolated from 1986 to 2021 in Korea (n = 188) were analyzed. Sequence alignments were carried out using Multiple alignment using Fast Fourier Transform of Geneious prime. The phylogenetic tree was generated using MEGA-11 (ver. 11.0.10) and Bayesian analysis was performed by BEAST v1.10.4. Selective pressure was analyzed via online server Datamonkey. Highlights and visualization of putative critical amino acid were conducted by using PyMol software (version 2.3). Results: Most (93.5%) belonged to the GI-19 lineage in Korea, and the GI-19 lineage was further divided into seven subgroups: KM91-like (Clade A and B), K40/09-like, QX-like (I-IV). Positive selection was identified at nine and six residues in S1 for KM91-like and QX-like IBVs, respectively. In addition, several positive selection sites of S1-NTD were indicated to have mutations at common locations even when new clades were generated. They were all located on the lateral surface of the quaternary structure of the S1 subunits in close proximity to the receptor-binding motif (RBM), putative RBM motif and neutralizing antigenic sites in S1. Conclusions: Our results suggest RBM surrounding sites in the S1 subunit of IBV are highly susceptible to mutation by selective pressure during evolution.

• Journal of Biosystems Engineering
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• 제41권1호
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• pp.60-65
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• 2016

Purpose: The necessity for precise manipulation of bioparticles has greatly increased in the fields of bioscience, biomedical, and environmental monitoring. Dielectrophoresis (DEP) is considered to be an ideal technique to manipulate bioparticles. The objective of this study is to develop a DEP microfluidic device that can trap fluorescent beads, which mimic bioparticles, at the low voltage and frequency of the sinusoidal signal supplied to the microfluidic device. Methods: A DEP microfluidic device, which is composed of polydimethylsiloxane (PDMS) channels and interdigitated electrode networks, is fabricated to trap fluorescent beads. The geometry of the interdigitated electrodes is determined through computational simulation. To determine the optimum voltage and frequency of the sinusoidal signal supplied to the device, the experiments of trapping beads are conducted at various combinations of voltage and frequency. The performance of the DEP microfluidic device is evaluated by investigating the correlation between fluorescent intensities and bead concentrations. Results: The optimum ratio of the widths between the negative and positive electrodes was 1:4 ($20:80{\mu}m$) at a gap of $20{\mu}m$ between the two electrodes. The DEP electrode networks were fabricated based on this geometry and used for the bead trapping experiments. The optimum voltage and frequency of the supplied signal for trapping fluorescent beads were 15 V and 5 kHz, respectively. The fluorescent intensity of the trapped beads increased linearly as the bead concentration increased. The coefficient of determination ($R^2$) between the fluorescent intensity and the bead concentration was 0.989. Conclusions: It is concluded that the microfluidic device developed in this study is promising for trapping bioparticles, such as a cell or virus, if they are conjugated to beads, and their concentration is quantified.

• Genomics & Informatics
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• 제14권3호
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• pp.78-84
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• 2016

Extranodal natural killer (NK)/T-cell lymphoma, nasal type (NKTCL), is a malignant disorder of cytotoxic lymphocytes of NK or T cells. It is an aggressive neoplasm with a very poor prognosis. Although extranodal NKTCL reportedly has a strong association with Epstein-Barr virus, the molecular pathogenesis of NKTCL has been unexplored. The recent technological advancements in next-generation sequencing (NGS) have made DNA sequencing cost- and time-effective, with more reliable results. Using the Ion Proton Comprehensive Cancer Panel, we sequenced 409 cancer-related genes to identify somatic mutations in five NKTCL tissue samples. The sequencing analysis detected 25 mutations in 21 genes. Among them, KMT2D, a histone modification-related gene, was the most frequently mutated gene (four of the five cases). This result was consistent with recent NGS studies that have suggested KMT2D as a novel driver gene in NKTCL. Mutations were also found in ARID1A, a chromatin remodeling gene, and TP53, which also recurred in recent NGS studies. We also found mutations in 18 novel candidate genes, with molecular functions that were potentially implicated in cancer development. We suggest that these genes may result in multiple oncogenic events and may be used as potential bio-markers of NKTCL in the future.

• Molecules and Cells
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• 제45권12호
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• pp.896-910
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• 2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible and potentially fatal virus. So far, most comprehensive analyses encompassing clinical and transcriptional manifestation have concentrated on the lungs. Here, we confirmed evident signs of viral infection in the lungs and spleen of SARS-CoV-2-infected K18-hACE2 mice, which replicate the phenotype and infection symptoms in hospitalized humans. Seven days post viral detection in organs, infected mice showed decreased vital signs, leading to death. Bronchopneumonia due to infiltration of leukocytes in the lungs and reduction in the spleen lymphocyte region were observed. Transcriptome profiling implicated the meticulous regulation of distress and recovery from cytokine-mediated immunity by distinct immune cell types in a time-dependent manner. In lungs, the chemokine-driven response to viral invasion was highly elevated at 2 days post infection (dpi). In late infection, diseased lungs, post the innate immune process, showed recovery signs. The spleen established an even more immediate line of defense than the lungs, and the cytokine expression profile dropped at 7 dpi. At 5 dpi, spleen samples diverged into two distinct groups with different transcriptome profile and pathophysiology. Inhibition of consecutive host cell viral entry and massive immunoglobulin production and proteolysis inhibition seemed that one group endeavored to survive, while the other group struggled with developmental regeneration against consistent viral intrusion through the replication cycle. Our results may contribute to improved understanding of the longitudinal response to viral infection and development of potential therapeutics for hospitalized patients affected by SARS-CoV-2.