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Anti-cancer activity of the ethylacetate fraction from Orostachys japonicus in A549 human lung cancer cells by induction of apoptosis and cell cycle arrest (인체 폐암 세포에 대한 와송 유래 에틸아세테이트 분획 생리 활성 물질의 세포사멸 유도 및 세포주기 억제 항암활성)

  • Kwon, Ji-Hye;Lee, Dong-Seok;Jung, Eun-Cheol;Kim, Hyeon-Mi;Kim, Su-Bin;Ryu, Deok-Seon
    • Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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    • v.7 no.1
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    • pp.395-405
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    • 2017
  • To confirm potential anti-cancer activities of ethylacetate (EtOAc) fraction from Orostachys japonicus on the A549 human lung cancer cells, this study examined. As a result of conducting MTS assay for measuring cell viability, the EtOAc fraction inhibited the proliferation of A549 cells in a dose-dependent manner. To investigate whether the inhibiting A549 cell viability was caused by apoptosis, this study analyzed chromatin condensation in A549 cells using DAPI staining. The morphological changes such as the formation of nuclear condensation were formed in a dose-dependent manner. Also, this study performed Annexin V-FITC staining for detecting phosphatidylinositol (PS). As a result of Annexin V-FITC staining to investigate level of early and late apoptosis, the apoptosis level treated with EtOAc fraction was higher than that of control. RT-PCR was performed to study the correlation between G2/M cell cycle arrest and cell cycle control genes. The anti-cancer activity of EtOAc fraction was accompanied by inhibition of CDK1, 4, cyclin B1 and D1 mRNA. This study also examined the expression of various marker proteins: p53, Bax, Bcl-2 and pro-caspase 3. Western blotting revealed that p53 and Bax proteins were up-regulated, and Bcl-2 and pro-caspase 3 proteins down-regulated in a time and dose-dependent manner.

A Genetically Encoded Biosensor for the Detection of Levulinic Acid

  • Tae Hyun Kim;Seung-Gyun Woo;Seong Keun Kim;Byeong Hyeon Yoo;Jonghyeok Shin;Eugene Rha;Soo Jung Kim;Kil Koang Kwon;Hyewon Lee;Haseong Kim;Hee-Taek Kim;Bong-Hyun Sung;Seung-Goo Lee;Dae-Hee Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.552-558
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    • 2023
  • Levulinic acid (LA) is a valuable chemical used in fuel additives, fragrances, and polymers. In this study, we proposed possible biosynthetic pathways for LA production from lignin and poly(ethylene terephthalate). We also created a genetically encoded biosensor responsive to LA, which can be used for screening and evolving the LA biosynthesis pathway genes, by employing an LvaR transcriptional regulator of Pseudomonas putida KT2440 to express a fluorescent reporter gene. The LvaR regulator senses LA as a cognate ligand. The LA biosensor was first examined in an Escherichia coli strain and was found to be non-functional. When the host of the LA biosensor was switched from E. coli to P. putida KT2440, the LA biosensor showed a linear correlation between fluorescence intensity and LA concentration in the range of 0.156-10 mM LA. In addition, we determined that 0.156 mM LA was the limit of LA detection in P. putida KT2440 harboring an LA-responsive biosensor. The maximal fluorescence increase was 12.3-fold in the presence of 10 mM LA compared to that in the absence of LA. The individual cell responses to LA concentrations reflected the population-averaged responses, which enabled high-throughput screening of enzymes and metabolic pathways involved in LA biosynthesis and sustainable production of LA in engineered microbes.

Lung Adenocarcinoma Mutation Hotspot in Koreans: Oncogenic Mutation Potential of the TP53 P72R Single Nucleotide Polymorphism (한국인의 폐선암 돌연변이 핫스팟: TP53 P72R Single Nucleotide Polymorphism의 발암성 돌연변이 가능성)

  • Jae Ha BAEK;Kyu Bong CHO
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.2
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    • pp.93-104
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    • 2023
  • This study aimed to identify new markers that cause lung adenocarcinoma by analyzing mutation hotspots for the top five genes with high mutation frequency in lung adenocarcinoma in Koreans by next generation sequencing (NGS) analysis. The association between TP53 mutation types and patterns with smoking, a major cause of lung cancer, was examined. The clinicopathological characteristics of lung adenocarcinoma patients with TP53 P72R SNPs were analyzed. In Korean lung adenocarcinoma cases, regardless of the smoking status, the TP53 P72R SNP was the most frequently occurring mutational hotspot, in which the nucleotide base was transversed from C to G, and the amino acid was substituted from proline to arginine at codon 72 of TP53. An analysis of the clinicopathological characteristics of lung adenocarcinoma cases with TP53 P72R SNP revealed no significant correlation with the patient's age, gender, smoking status, and tumor differentiation, but a significant correlation with low stage (P-value =0.026). This study confirmed an increase in TP53 rather than EGFR, which was reported as the most frequent mutations in lung adenocarcinoma in Koreans through NGS. Among them, TP53 P72R SNP is the most frequent regardless of smoking status.

Correlation of Protumor Effects of Leucine-Rich Repeat Kinase 2 with Interleukin-10 Expression in Lung Squamous Cell Carcinoma (폐 편평세포암종 내 Leucine-Rich Repeat Kinase 2 암촉진 효과와 Interleukin-10 발현과의 연관성)

  • Sung Won LEE;Sangwook PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.2
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    • pp.105-112
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    • 2023
  • Leucine-rich repeat kinase 2 (LRRK2) is known to play a crucial role in the pathophysiology of neurodegenerative disorders such as Parkinson's disease. LRRK2 is predominantly expressed in the lung as well as the brain. However, it is unclear whether LRRK2 expression correlates with the pathogenesis of lung squamous cell carcinoma (LUSC). This study analyzes the prognostic significance of LRRK2 in LUSC using the Kaplan-Meier plotter tool. High expression of LRRK2 is known to be associated with a bad prognosis in patients with LUSC. Patients with high LRRK2 expression, tumor mutational burden, high neoantigen load, and even gender correlation reportedly have the worse survival rates. In the gene expression profiling interactive analysis (GEPIA) database, the severity of pathogenesis in LUSC with high LRRK2 expression positively corresponds to a high expression of anti-inflammatory cytokines but not inflammatory cytokines. Similarly, the increased expression of interleukin (IL)10-related genes was shown to be significantly linked in LRRK2-high LUSC patients having a poor prognosis. Moreover, the tumor immune estimation resource (TIMER) database suggests that macrophages are one of the cellular sources of IL10 in LRRK2-high LUSC patients. Collectively, our results demonstrate that the postulated LRRK2-IL10 axis is a potential therapeutic target and prognostic biomarker for LUSC.

In-silico annotation of the chemical composition of Tibetan tea and its mechanism on antioxidant and lipid-lowering in mice

  • Ning Wang ;Linman Li ;Puyu Zhang;Muhammad Aamer Mehmood ;Chaohua Lan;Tian Gan ;Zaixin Li ;Zhi Zhang ;Kewei Xu ;Shan Mo ;Gang Xia ;Tao Wu ;Hui Zhu
    • Nutrition Research and Practice
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    • v.17 no.4
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    • pp.682-697
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    • 2023
  • BACKGROUND/OBJECTIVES: Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice. MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH2O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected. Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. RESULTS: The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. CONCLUSIONS: This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.

A Study on the Antibacterial Activity of Combined Administration of Jakyakgamcho-tang and Antibiotics Against MRSA (MRSA에 대한 작약감초탕과 항생제 병용투여의 항균활성에 관한 연구)

  • Dam Hee Kang;Ok Hwa Kang;Hee-Sung Chae;Dong Yeul Kwon
    • Korean Journal of Pharmacognosy
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    • v.54 no.2
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    • pp.72-79
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    • 2023
  • MRSA is Staphylococcus aureus resistant to β-lactam antibiotics, and is a worldwide infectious disease. Even with the discovery of new antibiotics, resistance develops rapidly, so new alternatives are needed. Jakyakgamcho-tang (JGT) is a combination of Jakyak and Gamcho, and has been mainly used as an antispasmodic and analgesic in oriental medicine. This study was conducted to find out whether there is an effect on MRSA in relation to the anti-inflammatory effect of JGT and the antibacterial effect of Jakyak and Gamcho found in previous studies. In this study, in order to investigate the antibacterial activity of JGT and the combined effect of existing antibiotics, after extracting JGT with 70% EtoH, the disc diffusion method, minimum inhibitory concentration (MIC), drug combination effect (FICI), and time-kill analysis (Time-kill assay), metabolic inhibition, Western blot and qRT-PCR analysis were used to confirm the antibacterial activity mechanism of MRSA of JGT. As a result of the experiment, all of MRSA showed antibacterial activity in JGT's disc diffusion method, and the MIC was 250-1000 ㎍/mL. When existing antibiotics and JGT were combined with drugs, most had synergy or partial synergy. In addition, it was confirmed that the degree of bacterial growth was suppressed over time when simultaneous administration for 24 hours. JGT showed a synergistic effect when administered together with the ATPase-inhibitor DCCD, suggesting that it affected the inhibition of ATPase. As a result of observing the expression of PBP2a, and hla protein in the JGT-treated group and the untreated control group through wstern blot, it was confirmed that the protein expression of the JGT-treated group was significantly suppressed, and the expression levels of mecA, mecR1 and hla genes were also suppressed during JGT treatment. was observed by qRT-PCR. Combining the results of the experiment, it can be seen that JGT has antibacterial activity in MRSA, and when combined with existing antibiotics, the effect was increased compared to treatment with the drug alone. This suggests that JGT can be an alternative to treatment for antibiotic resistance of MRSA.

Whole-genome resequencing reveals domestication and signatures of selection in Ujimqin, Sunit, and Wu Ranke Mongolian sheep breeds

  • Wang, Hanning;Zhong, Liang;Dong, Yanbing;Meng, Lingbo;Ji, Cheng;Luo, Hui;Fu, Mengrong;Qi, Zhi;Mi, Lan
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1303-1313
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    • 2022
  • Objective: The current study aimed to perform whole-genome resequencing of Chinese indigenous Mongolian sheep breeds including Ujimqin, Sunit, and Wu Ranke sheep breeds (UJMQ, SNT, WRK) and deeply analyze genetic variation, population structure, domestication, and selection for domestication traits among these Mongolian sheep breeds. Methods: Blood samples were collected from a total of 60 individuals comprising 20 WRK, 20 UJMQ, and 20 SNT. For genome sequencing, about 1.5 ㎍ of genomic DNA was used for library construction with an insert size of about 350 bp. Pair-end sequencing were performed on Illumina NovaSeq platform, with the read length of 150 bp at each end. We then investigated the domestication and signatures of selection in these sheep breeds. Results: According to the population and demographic analyses, WRK and SNT populations were very similar, which were different from UJMQ populations. Genome wide association study identified 468 and 779 significant loci from SNT vs UJMQ, and UJMQ vs WRK, respectively. However, only 3 loci were identified from SNT vs WRK. Genomic comparison and selective sweep analysis among these sheep breeds suggested that genes associated with regulation of secretion, metabolic pathways including estrogen metabolism and amino acid metabolism, and neuron development have undergone strong selection during domestication. Conclusion: Our findings will facilitate the understanding of Chinese indigenous Mongolian sheep breeds domestication and selection for complex traits and provide a valuable genomic resource for future studies of sheep and other domestic animal breeding.

Effects of different cardiorespiratory fitness response to exercise training on cardiovascular disease and adipocytokine in abdominal obesity women (운동 트레이닝을 통한 심폐체력 반응의 차이가 복부비만 여성의 심혈관계 위험요인과 아디포싸이토카인에 미치는 영향)

  • Park, S.H.
    • Exercise Science
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    • v.21 no.1
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    • pp.111-120
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    • 2012
  • The purpose of the study was to assess the effects of different cardiorespiratory fitness response to exercise training on cardiovascular disease and adipocytokine release and gene expression in abdominal obesity women. forty eight middle-age women were divided to a exercise-responder (n=34) group (ERG) and a exercise-nonresponder (n=14) group (ENRG) based on cardiorespiratory fitness after exercise training (12weeks, 1200 kcal/week, moderate-vigorous intensity, walking and jogging). Cardiorespiratory fitness was measured using maximal oxygen uptake with metabolic gas analysis and body composition was determined by bioelectrical impedance. We measured lipid and glucose profiles, blood adipocytokines and adipocytokine genes expression in adipose tissue. Waist girth (p=0.040), and %body fat (p=0.031) were significantly decreased in ERG than ENRG. Triglyceride (p=0.023) and systolic blood pressure (0.046) were significantly decreased in ERG than ENRG. Blood leptin (p=0.022) was significantly decrease in ERG than ENRG but leptin gene expression was decreased both groups (p<0.001). These results show that exercise-responder group improved cardiovascular disease risk factors and adipocytokine more than exercise-nonresponder group after exercise training. Moreover, exercise-nonresponders group show that obese indices and blood leptin and leptin gene expression in adipose tissue were decreased despite the failure to improve fitness. Therefore, regular exercise training seems to give health benefits although the failure to improve fitness.

Present and Prospect of Plant Genomics in Japan (일본의 식물유전체 연구현황 및 전망)

  • Yoon, Ung-Han;Lee, Jeonghwa;Lee, Gang-Seob;Kim, Young-Mi;Ji, Hyeon-So;Kim, Tae-Ho
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.560-569
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    • 2011
  • In Japan, plant genomics research is mainly leaded by the national research institutes. The various structural studies such as rice genome has allowed researchers to analyze useful traits, and to focus their commercialization. With aims to facilitate structural and functional study in plant genome, NIAS (National Institute of Agrobiological Sciences) constructed Poaceae genome DB and RIKEN (Rikagaku Kenkyusho) built DB for Arabidopsis genome and plant full-length cDNA. NIG (National Institute of Genetics) constructed a national biological resources DB (National Bio Resource Project). This compiling DB provides a variety of genome-related research materials for researchers in the field. Recently, as an effort to resolve global issues of food supply and environmental problems, New Agri-genome Project has been performed aiming to develop an innovative agricultural technologies for the quantity, disease resistance and identifying useful genes related to environmental problems. In addition, in order to improve agricultural productivity in developing countries, JIRCAS assisted technical supports for the plant genome research and developed NERICA rice, which is suitable for African area. Such these approaches are expected to contribute to solving the global issues about food, energy and environment in the world.

Characterization of a Gamma Radiation-Induced Salt-Tolerant Silage Maize Mutant (방사선 유도 내염성 증진 사료용 옥수수 돌연변이체 특성 분석)

  • Cho, Chuloh;Kim, Kyung Hwa;Choi, Man-Soo;Chun, Jaebuhm;Seo, Mi-Suk;Jeong, Namhee;Jin, Mina;Son, Beom-Young;Kim, Dool-Yi
    • Korean Journal of Breeding Science
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    • v.51 no.4
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    • pp.318-325
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    • 2019
  • Salt stress is a significant factor limiting growth and productivity in crops. However, little is known about the response and resistance mechanism to salt stress in maize. The objective of this research was to develop an enhanced salt-tolerant silage maize by mutagenesis with gamma radiation. To generate gamma radiation-induced salt-tolerant silage maize, we irradiated a KS140 inbred line with 100 Gy gamma rays. Salt tolerance was determined by evaluating plant growth, morphological changes, and gene expression under NaCl stress. We screened 10 salt-tolerant maize inbred lines from 2,248 M2 mutant populations and selected a line showing better growth under salt stress conditions. The selected 140RS516 mutant exhibited improved seed germination and plant growth when compared with the wild-type under salt stress conditions. Enhanced salt tolerance of the 140RS516 mutant was attributed to higher stomatal conductance and proline content. Using whole-genome re-sequencing analysis, a total of 328 single nucleotide polymorphisms and insertions or deletions were identified in the 140RS516 mutant. We found that the expression of the genes involved in salt stress tolerance, ABP9, CIPK21, and CIPK31, was increased by salt stress in the 140RS516 mutant. Our results suggest that the 140RS516 mutant induced by gamma rays could be a good material for developing cultivars with salt tolerance in maize.