• Title/Summary/Keyword: bacterial inhibition

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Inhibition of Lipopolysaccharide-Inducible Nitric Oxide Synthase, $TNF-{\alpha}$, $IL-1{\beta}$ and COX-2 Expression by Flower and Whole Plant of Lonicera japonica (금은화(金銀花) 및 금은화전초(金銀花全草)가 Raw 264.7 cell에서 LPS로 유도된 NO의 생성, iNOS, COX-2 및 cytokine에 미치는 영향)

  • Lee, Dong-Eun;Lee, Jae-Ryung;Kim, Young-Woo;Kwon, Young-Kyu;Byun, Sung-Hui;Shin, Sang-Woo;Suh, Seong-Il;Kwon, Taeg-Kyu;Byun, Joon-Seok;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.481-489
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    • 2005
  • Lonicerae Flos has antibacterial effects against Staphylococcus aureus, streptococci, pneumococci, Bacillus dysenterii, Salmonella typhi, and paratyphoid. It is an antiviral agent. The herb has a cytoprotective effect against $CCl_{4}-induced$ hepatic injury. It has antilipemic action, interfering with lipid absorption from the gut. Nowadays this herb is used mainly in the treatment of upper respiratory infections, such as tonsillitis and acute laryngitis. It is also used in the treatment of skin suppurations, such as carbuncles, and to treat viral conjunctivitis, influenza, pneumonia, and mastitis. Lonicerae Flos is dried flower buds of Lonicera japonica, L. hypoglauca, L. confusa, or L. dasystyla. But, for the most part, we use whole plant of Lonicera japonica, as a flower bud of it. And, little is known of the original copy of effects of whole plant, except for the 'Bon-Cho-Gang-Mok', which is written the effects of flower of Lonicera japonica are equal to effects of leaves and branch of it. The present study was conducted to evaluate the effect of flower and whole plant of Lonicera japonica on the regulatory mechanism of cytokines, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) for the immunological activities in Raw 264.7 cells. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, flower and whole plant of Lonicera japonica water extracts inhibited nitric oxide production in a dose-dependent manner and abrogated iNOS and COX-2. Flower and whole plant of Lonicera japonica water extract did not affect on cell viability. To investigate the mechanism by which flower and whole plant of Lonicera japonica water extract inhibits iNOS and COX-2 gene expression, we examined the on phosphorylation of inhibitor ${\kappa}B{\alpha}$ and assessed production of $TNF-{\alpha}$, $interleukin-1{\beta}$ $(IL-1{\beta})$ and interleukin-6 (IL-6). Results provided evidence that flower and whole plant of Lonicera japonica inhibited the production of $IL-1{\beta}$, IL-6 and activated the phosphorylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with LPS. These findings suggest that flower and whole plant of Lonicera japonica can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections, respectively.

Apoptotic Effect of Extract from Artemisia annua Linné by Akt/mTOR/GSK-3β Signal Pathway in Hep3B Human Hepatoma Cells (Hep3B 간암세포에서 개똥쑥추출물로부터 Akt-mTOR-GSK3β 신호경로에 의한 apoptosis 효과)

  • Kim, Eun Ji;Kim, Guen Tae;Kim, Bo Min;Lim, Eun Gyeong;Ha, Sung Ho;Kim, Sang-Yong;Kim, Young Min
    • Journal of Life Science
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    • v.26 no.7
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    • pp.764-771
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    • 2016
  • Extracts from Artemisia annua Linné (AAE) have been known to possess various functions, including anti-bacterial, anti-virus, and anti-oxidant effects. However, the mechanism of those effects of AAE is not well-known. The aim of this study was to analyze the inhibitory effects of AAE on cell proliferation of the human hepatoma cell line (Hep3B) and to examine its effects on apoptosis. Activation by phosphorylation of Akt is cell proliferation through the phosphorylation of TSC2, mTOR, and GSK-3β. We suggested that AAE may exert cancer cell apoptosis through Akt/mTOR/GSK-3β signal pathways and mitochondria-mediated apoptotic proteins. For this, we examined the effects of extracts of AAE on cell proliferation according to treatment concentration. Treatment with AAE not only reduced cell viability, but also resulted in the induced release of lactate dehydrogenase (LDH). These results were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a lactate dehydrogenase (LDH) assay. Furthermore, we determined the effects of apoptosis through Hoechst 33342 staining, annexinⅤ-propidium iodide (PI) staining, 5,5′, 6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide (JC-1) staining, and Western blotting. Our study showed that the treatment of liver cancer cells with AAE resulted in the inhibition of Akt, TSC2, GSK-3β-phosphorylated, Bcl-2, and pro-caspase 3 and the activation of Bim, Bax, Bak, and cleaved PARP expressions. These results indicate that AAE induced apoptosis by means of a mitochondrial event through the regulate of Akt/mTOR/GSK-3β signaling pathways.

Production of Antimicrobial Compounds and Cloning of a dctA Gene Related Uptake of Organic Acids from a Biocontrol Bacterium Pseudomonas Chlororaphis O6 (생물적 방제균 Pseudomonas chlororaphis O6의 길항 물질 생산 및 유기산 흡수에 관련된 dctA 유전자의 클로닝)

  • Han, Song-Hee;Nam, Hyo-Song;Kang, Beom-Ryong;Kim, Kil-Yong;Koo, Bon-Sung;Cho, Baik-Ho;Kim, Young-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.36 no.3
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    • pp.134-144
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    • 2003
  • A rhizobacterium Pseudomonas cholororaphis O6 produced several secondary metabolites, such as phenazines, protease, and HCN that may be involved in inhibition of the growth of phytopathogenic fungi. In field study, P. chlororaphis O6 treatment on wheat seed suppressed root rot disease caused by Fusarium culmorum. The major organic acids of cucumber root exudates were fumaric acid, malic acid, benzoic acid, and succinic acid. Glucose and fructose were major monosaccharides in cucumber root exudates. The total amount of organic acids was ten times higher than that of the sugars. P. chlororaphis O6 grew well on cucumber root exudates. The dctA gene of P. chlororaphis O6 consisted of a 1,335 bp open reading frame with a deduced amino acid sequence of 444 residues, corresponding to a molecular size of about 47 kD and pI 8.2. The deduced dctA sequence has ten putative transmembrane domains, as expected of a membrane-embedded protein. Our results indicated that organic acids in cucumber root exudates may play an important role in providing nutrient source for root colonization of biological control bacteria, and the dctA gene of P. chlororaphis O6 may be an important bacterial trait that is involved in utilization of root exudates.

Characterization of Agarase from a Marine Bacterium Agarivorans sp. BK-1 (해양세균 Agarivorans sp. BK-1의 분리 및 β-아가라제의 특성 규명)

  • Ahn, Byeong-Ki;Min, Kyung-Cheol;Lee, Dong-Geun;Kim, Andre;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1173-1178
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    • 2019
  • The purpose of this study was to isolate an agar-degrading marine bacterium and characterize its agarase. Bacterium BK-1, from Gwanganri Beach at Busan, Korea, was isolated on Marine 2216 agar medium and identified as Agarivorans sp. BK-1 by 16S rRNA gene sequencing. The extracellular agarase, characterized after dialysis of culture broth, showed maximum activity at pH 6.0 and $50^{\circ}C$ in 20 mM Tris-HCl buffer. Relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 67, 93, 97, 100, 58, and 52%, respectively. Relative activities at pH 5, 6, 7, and 8 were 59, 100, 95, and 91%, respectively. More than 90% of the activity remained after a 2 hr exposure to 20, 30, or $40^{\circ}C$; about 60% of the activity remained after a 2 hr exposure to $50^{\circ}C$. Almost all activity was lost after exposure to 60 or $70^{\circ}C$ for 30 min. Zymography revealed three agarases with molecular weights of 110, 90, and 55 kDa. Agarose was degraded to neoagarobiose (46.8%), neoagarotetraose (39.7%), and neoagarohexaose (13.5%), confirming the agarase of Agarivorans sp. BK-1 as a ${\beta}$-agarase. The neoagarooligosaccharides generated by this agarase could be used for moisturizing, bacterial growth inhibition, skin whitening, food treatments, cosmetics, and delaying starch degradation.

Effect of Sterilization Conditions on Microbial Reduction in Cleaning Tools (살균 조건이 세척 도구 중 미생물 저감화에 미치는 영향)

  • Im, Ji-Yu;Kim, Chae-Young;Kim, Eun-yeong;Kim, Min-jin;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.37 no.5
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    • pp.310-316
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    • 2022
  • In this study, we compared the microbial reduction effects of drying, hot water, and microwave sterilization in scourers and dishcloths to suggest a most suitable sterilization method. Three scourer types (silver, copper, and mesh) were used, and three dishcloth types (silver, bamboo, and cotton) were used. Drying time dependent reduction in Escherichia coli was high in silver and copper scourers, but minimal bacterial reduction was obtained against Bacillus cereus in all scourers and dishcloths. In scourers, E. coli was not detected after ≥30 s of hot water sterilization at 77℃, and B. cereus was not detected after ≥60 s of hot water sterilization at 100℃. In dishcloths, E. coli was not detected after hot water sterilization at 77℃ for ≥30 s, but B. cereus was detected after hot water sterilization at 100℃ for ≥60 s. In scourers, E. coli was not detected after microwave sterilization at 700 W for 3 min, but B. cereus was detected. In dishcloths, E. coli was not detected after microwave sterilization with 700 W for ≥1 min, but B. cereus was detected in the cotton dishcloth even after sterilization for 3 min. In conclusion, the use of antimicrobial scourers (silver and copper) and dishcloths (silver and bamboo) are not sufficient to reduce the microbial contamination. The guideline provided by the Ministry of Food and Drug Safety suggesting dishcloth sterilization via hot water at 100℃ for 30 s was also found to be insufficient. Based on our research, we suggest that the most effective methods of microbial management are submerging scourers in hot water at 100℃ for ≥1 min, and sterilizing dishcloths for ≥3 min using a 700 W microwave.

Isolation of Agarivorans sp. JS-1 and Characterization of Its β-Agarase (한천분해세균 Agarivorans sp. JS-1의 분리 및 β-아가라제의 특성 규명)

  • Jin Sun Kim;Dong-Geun Lee;Go-Wun Yeo;Min-Joo Park;Sang-Hyeon Lee
    • Journal of Life Science
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    • v.33 no.4
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    • pp.357-362
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    • 2023
  • This report looks at an agar-degrading marine bacterium and characterization of its agarase. Agar-degrading marine bacterium JS-1 was isolated with Marine agar 2216 media from seawater from the seashore of Sojuk-do, Changwon in Gyeongnam Province, Korea. The agar-degrading bacterium was named as Agarivorans sp. JS-1 by phylogenetic analysis based on 16S rRNA gene sequencing. The extracellular agarase was prepared from the culture media of Agarivorans sp. JS-1 and used for characterization. Relative activities at 20℃, 30℃, 35℃, 40℃, 45℃, 50℃, 55℃, and 60℃ were 70%, 74%, 78%, 83%, 87%, 100%, 74%, and 66%, respectively. Relative activities at pH 5, 6, 7, and 8 were 91%, 100%, 90%, and 89%, respectively. Its extracellular agarase showed maximum activity (207 units/l) at pH 6.0 and 50℃ in 20 mM Tris-HCl buffer. The residual activity after heat treatment at 20℃, 30℃, and 50℃ for 30 minutes was 90%, 70%, and 50% or more, respectively. After a 2-hour heat treatment at 20℃, 30℃, 35℃, 40℃, and 45℃, the residual activity was 80%, 68%, 65%, 63%, and 57%, respectively. At 50℃ and above, after heat treatment for 30 minutes, the residual activity was below 60%. Thin layer chromatography analysis suggested that Agarivorans sp. JS-1 produces extracellular β-agarases as they hydrolyze agarose to produce neoagarooligosaccharides such as neoagarohexaose (20.6%), neoagarotetraose (58.5%), and neoagarobiose (20.9%). Agarivorans sp. JS-1 and its thermotolerant β-agarase would be useful in the production of neoagarooligosaccharides, showing functional activity such as inhibition of bacterial growth and delay of starch degradation.

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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