• Title/Summary/Keyword: Resistant Genes

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Comparative Analysis on Antioxidant Activity in Various Human Skin Fibroblasts and Mesenchymal Stem Cells (사람의 피부 섬유아세포 및 중간엽 줄기세포에서 항산화 활성의 반응에 대한 비교 분석)

  • Kong, Ji-Weon;Park, Ryeok;Park, Joon-Woo;Lee, Joo-Yeong;Choi, Yeon-Joo;Moon, Sun-Ha;Kim, Hyeon-Ji;Jeon, Byeong-Gyun
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.394-406
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    • 2019
  • The cellular senescence may be due to damage by the reactive oxygen species (ROS). This study has compared the antioxidant activity in the human cell lines of various origins, including 10S and 50S-derived normal skin fibroblasts, and 10S bone marrow, dental tissue and adipose-derived adult stem cells. After being exposed to $H_2O_2$, half inhibitory concentration ($IC_{50}$) values by cytotoxicity assay was significantly (P<0.05) lower in 50S-derived skin fibroblasts, than in 10S-derived skin fibroblasts and various adult stem cell lines. The cell population doubling time (PDT) and the cell frequency with high senescence associated-${\beta}$-galactose activity were remarkably increased in 50S-derived fibroblasts exposed to 50 ppm $H_2O_2$ for 7 days, than those of 10S-derived fibroblasts and various adult stem cell lines. Further, the expression level of antioxidant-related genes, glutathione peroxidase (GPX) and catalase (CAT), was investigated in 10S and 50S-derived skin fibroblasts, and 10S-derived various adult stem cells by reverse transcription polymerase chain reaction (RT-PCR). The expression level of GPX was higher in most of cell lines, compared to CAT, and a significantly (P<0.05) higher expression level of GPX was observed in 10S-derived skin fibroblasts and adult stem cell lines, compared to 50S-derived skin fibroblasts. We concluded that old-aged skin fibroblasts seemed to be less resistant against ROS than young-aged skin fibroblasts and adult stem cells.

Inhibitory Effects of Locusta migratoria Ethanol Extracts on RANKL-induced Osteoclast Differentiation (RANKL 유도된 파골세포 분화에 대한 풀무치 에탄올 추출물의 분화 억제 효과)

  • Baek, Minhee;Seo, Minchul;Lee, Joon Ha;Lee, Hwa Jeong;Kim, In-Woo;Kim, Sun Young;Kim, Mi-Ae;Kim, Sunghyun;Hwang, Jae-Sam
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1104-1110
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    • 2019
  • Recently, there has been an increase in the elderly population of the world. Consequently, bone metabolic diseases such as osteoporosis are emerging as a social problem. Osteoclasts play a role in bone resorption, and osteoporosis is induced when bone resorption occurs excessively. Because currently used bone resorption inhibitors may cause side effects when used for a long period of time, it is necessary to develop a new material that effectively inhibits osteoclast differentiation. This study aimed to confirm the inhibitory effect of ethanol extract of Locusta migratoria on RANKL-induced osteoclast differentiation and its mechanism. The toxicity and proliferation effects of LME on RAW264.7 osteoclasts were measured by an MTS assay. There was no cytotoxicity or proliferation when the osteoclasts were treated with up to $2,000{\mu}g/ml$ of LME. In order to confirm the effect of LME on the differentiation of osteoclasts, osteoclasts were treated with RANKL alone or with LME for 3 days. As a result of a TRAP (tartrate-resistant acid phosphatase) assay, the increasing osteoclast differentiation by RANKL decreased in a concentration-dependent manner with the treatment of LME. In addition, LME suppressed the expression of differentiation-related marker genes (TRAP, RANK, NFATc1, and CK) and proteins (NFATc1 and c-Src) that had been increased by RANKL. Also, LME influenced the $NF-{\kappa}B$, ERK and JNK signaling pathways, resulting in the inhibition of osteoclast differentiation. These results suggest that LME may be used as a novel functional material for the prevention and treatment of osteoporosis by playing a role in inhibiting bone absorption.

Change in the Sensitivity to Propiconazole of Fusarium graminearum Species Complex Causing Head Blight of Barley and Wheat in Jeolla Province (전남북 지역 맥류 붉은곰팡이병균의 Propiconazole 약제에 대한 감수성 변화)

  • Jiseon Baek;Ju-Young Nah;Mi-Jeong Lee;Su-Bin Lim;Jung-Hye Choi;Ja Yeong Jang;Theresa Lee;Hyo-Won Choi;Jeomsoon Kim
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.281-289
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    • 2022
  • Fusarium head blight is an important disease of small grains. It is mainly caused by members of the Fusarium graminearum species complex (FGSC). Barley and wheat growers spray fungicides, especially demethylation-inhibitor fungicides, to suppress the disease. The objective of this study was to examine the changes in the sensitivity of the FGSC population to the triazole fungicide, propiconazole. A total of 124 and 350 isolates of FGSC were obtained from barley and wheat in Jeolla Province during 2010-2016 and 2020-2021, respectively. The species identity and trichothecene chemotypes of the FGSC isolates were determined based on polymerase chain reaction assays targeting translation elongation factor 1-alpha and TRI12 genes, respectively. Sensitivity to propiconazole was determined based on the effective concentration that reduced 50% of the mycelial growth (EC50) using the agar dilution method. Of all isolates, F. asiaticum with the nivalenol chemotype was the most common (83.9% in 2010-2016 and 96.0% in 2020-2021), followed by F. asiaticum with the 3-acetyl deoxynivalenol chemotype (12.1% in 2010-2016 and 2.9% in 2020-2021). The EC50 values of the isolates collected in 2010-2016 and 2020-2021 ranged from 0.0180 to 11.0166 ㎍/mL and 1.3104 to 17.9587 ㎍/mL, respectively. The mean EC50 value of the isolates increased from 3.8648 ㎍/mL in 2010-2016 to 5.9635 ㎍/mL in 2020-2021. The baseline resistance to propiconazole was determined to be 7 ㎍/mL, based on the EC50 value of isolates collected in 2010-2016, and the ratio of resistant isolates increased from 9.7% in 2010-2016 to 28.6% in 2020-2021.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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