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Development of Cleaved Amplified Polymorphic Sequence Markers for the Identification of Lentinula edodes Cultivars Sanmaru 1ho and Chunjang 3ho (표고버섯 품종 산마루1호, 천장3호를 구분할 수 있는 CAPS Marker 개발)

  • Moon, Suyun;Lee, Hwa-Yong;Kim, Myungkil;Ka, Kang-Hyeon;Ko, Han Kyu;Chung, Jong-Wook;Koo, Chang-Duck;Ryu, Hojin
    • The Korean Journal of Mycology
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    • v.45 no.2
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    • pp.114-120
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    • 2017
  • Lentinula edodes is an edible mushroom that is mainly cultivated in Asian countries. Recently, new cultivars of this mushroom have been developed in Korea; variety protection is very important, so the development of efficient molecular markers that can distinguish each variety is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanmaru 1ho and Chunjang 3ho). These markers were developed from whole genomic sequencing data from L. edodes monokaryon strain B17 and resequencing data from 10 dikaryon strains. A single nucleotide polymorphism changed in scaffold 9 POS 1630048 in Sanmaru 1ho($G{\rightarrow}T$), and in scaffold 13 POS 920681 in Chunjang 3ho ($G{\rightarrow}A$). The restriction enzymes TspR I and Xho I distinguished Sanmaru 1ho and Chunjang 3ho, respectively, from other strains. Thus, we developed 2 CAPS markers for the identification of the L. edodes cultivars Sanmaru 1ho and Chunjang 3ho.

Effect of Allium cepa (red) and Angelica gigas Nakai on Hair-growth Promotion in C57BL/6 Mice (C57BL/6 마우스에서 모발성장 촉진에 대한 Allium cepa (red)와 Angelica gigas Nakai의 효과)

  • Lee, Jin Young;Dong, Jae Kyung;Kang, Jae Seon
    • Journal of Life Science
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    • v.30 no.11
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    • pp.990-998
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    • 2020
  • This study was conducted to evaluate the hair-growth promotion effect in C57BL/6 mice of a new substance mixed with Allium cepa (red)-bioconversion extract and Angelica gigas Nakai. The ethanol extract of Allium cepa (red) was bioconverted through the use of the Bacillus subtilis KJ-3 (BS3) strain, which was named Red-BCQ. The quercetin content of Red-BCQ increased by about 7.4-fold after bioconversion. Angelica gigas Nakai extract (Agnex) contains a large amount of coumarins such as decursin (D) and decursinol angelate (DA). A 1 mg portion of Agnex contained 0.4146 mg of D and 0.3659 mg of DA. Minoxidil has been known to promote hair growth. In this study, the hair-growth promotion effects of Red-BCQ, Agnex, and a mixture of both Red-BCQ and Agnex were compared with 5% minoxidil. Twenty-five mice were divided into five experimental groups including saline (CON), 5% minoxidil (PCON), Red-BCQ (RA), Agnex (AG), and a Red-BCQ-Agnex mixture (RAG)-treated group. Samples were administered orally once a day at a fixed time for 4 weeks. Hair growth was monitored by photograph at 7, 14, 21, and 28 days. We also observed 5α-reductase, alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GT), insulin-like growth factor (IGF)-1, transforming growth factor (TGF)-β1, antioxidant enzyme, and the hair follicles of the skin tissue. In all the results, the RAG-administered group showed greater antioxidative and hair-growth promotion effects than the other groups. These data suggest that RAG has potent stimulating activity on hair growth in C57BL/6 mice.

Anti-Toxoplasmosis Effect of the Herbal Extracts Plantago asiatica L. (차전초(Plantago asiatica L.)의 항톡소포자충증 효과)

  • Hong, Sunhwa;Kim, Okjin
    • Korean Journal of Plant Resources
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    • v.29 no.1
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    • pp.26-31
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    • 2016
  • Toxoplasmosis is an important cause of foodborne, inflammatory illnesses, as well as congenital abnormalities. Currently available therapies are ineffective for persistent chronic disease and congenital toxoplasmosis or have severe side effects which may result in life-threatening complications. There is an urgent need for safe and effective therapies to eliminate or treat this cosmopolitan infectious disease. The aim of this study was to investigate the in vitro anti-Toxoplasma activities of Plantago asiatica L., one of the herbal extracts, using tachyzoit of T. gondii RH strain infected HeLa cells. As the results, the selectivity of Plantago asiatica L. extract was 4.5, which was higher than Sulfadiazine selectivity (0.4). Also, we perfomed the cell proliferation inhibition test and the morphological study to evaluate the anti-T. gondii activity of Plantago asiatica L. extract with HeLa cells. As the results, the inhibition rate of the Plantago asiatica L. extract was high inhibition rate. This indicates that the Plantago asiatica L. extract may be used for new anti-T. gondii agent.

Antibacterial Activity and Probiotic Potential of Lactobacillus plantarum HKN01: A New Insight into the Morphological Changes of Antibacterial Compound-Treated Escherichia coli by Electron Microscopy

  • Sharafi, Hakimeh;Maleki, Hadi;Ahmadian, Gholamreza;Zahiri, Hossein Shahbani;Sajedinejad, Neda;Houshmand, Behzad;Vali, Hojatollah;Noghabi, Kambiz Akbari
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.225-236
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    • 2013
  • Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.

Isolation, Purification, and Characterization of Five Active Diketopiperazine Derivatives from Endophytic Streptomyces SUK 25 with Antimicrobial and Cytotoxic Activities

  • Alshaibani, Muhanna M.;MohamadZin, Noraziah;Jalil, Juriyati;Sidik, Nik Marzuki;Ahmad, Siti Junaidah;Kamal, Nurkhalida;Edrada-Ebel, RuAngelie
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1249-1256
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    • 2017
  • In our search for new sources of bioactive secondary metabolites from Streptomyces sp., the ethyl acetate extracts from endophytic Streptomyces SUK 25 afforded five active diketopiperazine (DKP) compounds. The aim of this study was to characterize the bioactive compounds isolated from endophytic Streptomyces SUK 25 and evaluate their bioactivity against multiple drug resistance (MDR) bacteria such as Enterococcus raffinosus, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Enterobacter spp., and their cytotoxic activities against the human hepatoma (HepaRG) cell line. The production of secondary metabolites by this strain was optimized through Thornton's medium. Isolation, purification, and identification of the bioactive compounds were carried out using high-performance liquid chromatography, high-resolution mass liquid chromatography-mass spectrometry, Fourier transform infrared spectroscopy, and nuclear magnetic resonance, and cryopreserved HepaRG cells were selected to test the cytotoxicity. The results showed that endophytic Streptomyces SUK 25 produces four active DKP compounds and an acetamide derivative, which were elucidated as $cyclo-({\text\tiny{L}}-Val-{\text\tiny{L}}-Pro)$, $cyclo-({\text\tiny{L}}-Leu-{\text\tiny{L}}-Pro)$, $cyclo-({\text\tiny{L}}-Phe-{\text\tiny{L}}-Pro)$, $cyclo-({\text\tiny{L}}-Val-{\text\tiny{L}}-Phe)$, and N-(7-hydroxy-6-methyl-octyl)-acetamide. These active compounds exhibited activity against methicillin-resistant S. aureus ATCC 43300 and Enterococcus raffinosus, with low toxicity against human hepatoma HepaRG cells. Endophytic Streptomyces SUK 25 has the ability to produce DKP derivatives biologically active against some MDR bacteria with relatively low toxicity against HepaRG cells line.

Protection of Infection and Eradication Activity of Culture Product by Pediococcus pentosaceus CBT SL4 Showing Antimicrobial Activity against Helicobacter pylori (Helicobacter pylori에 대한 항균활성을 나타내는 Pediococcus pentosaceus CBT SL4 배양물의 감염방어 및 제균활성)

  • Hong, Un-Pyo;Chung, Myung-June;Kim, Soo-Dong;Oh, Eun-Taex;So, Jae-Seong;Chung, Chung-Il
    • Korean Journal of Food Science and Technology
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    • v.36 no.5
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    • pp.779-783
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    • 2004
  • New food ingredient was developed to eradicate and protect against re-infection of Helicobacter pylori in fermentation broth of lactic acid bacteria (LAB) showing antimicrobial activity against pathogenic microorganisms such as H. pylori and Listeria monocytogenes. LAB strain CBT SL4 was identified as Pediococcus pentosaceus by 16S rDNA sequencing and its culture broth showed antimicrobial activity of 800 AU/mL against H. pylori in optimized fermentation process. Using thin layer concentration system and spray-typed fluid bed drier system, concentrated powder product showing activity of 12,800 AU/g was harvested. Product showed eradication and protection activities against H. pylori infection on feeding test (50 AU/day) using Mongolian gerbil infection model. After 4 weeks therapy of 8,000 AU/day, ${\Delta}13CO_2$ level (DOB30) decreased about 40% in urea breath test on patient with H. pylori infection. Result show concentrated culture product of P. pentosaceus CBT SL4 has eradicating and protecting activities against H. pylori infection and can be used as food-active ingredient for prevention of gastric and duodenum ulcer caused by H. pylori.

A Stability Analysis of Geosynthetics Reinforced Soil Slopes II - Evaluation of Required Reinforcement Tensile Force - (토목섬유 보강 성토사면의 안정해석 II. - 소요 보강재 인장력 평가 -)

  • Kim Kyeong-Mo;Kim Hong-Taek;Lee Eun-Soo;Kim Young-Yoon;Ahn Kwang-Kuk
    • Journal of the Korean Geotechnical Society
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    • v.21 no.8
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    • pp.107-116
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    • 2005
  • Generally, when the stability of the geosynthetic reinforced soil slopes is analyzed, the required tensile forces of each reinforcement layers are calculated from total reinforcement forces which are necessary to retain the equilibrium state of slip mass in which the slip surfaces are assumed to be a linear or bilinear. It is assumed that the reinforcement forces are increased or constant with depth. However, according to the instrumented field data and laboratory model test results, the maximum tensile strain of reinforcement in a reinforced soil slope is developed in a certain elevation, not a bottom of the slope. In the concept of reinforced soil, postulated failure surfaces are the traces of the position in which the reinforcement tensile forces are maximum in the layer, and the reinforcement tensile forces are related to the stress state on the postulated failure surface. Therefore, in this study, based on the distribution of normal stress on the slip surface, a new method for the evaluation of required tensile forces is suggested and a number of the instrumented field data are analyzed by newly suggested method. As a result, it is shown that the newly suggested method produces relatively accurate reinforcement tension forces.

Optimization of Anion-exchange Chromatography for the Separation of Agarase from Culture Broth of Pseudoalteromonas sp. (Pseudoalteromonas sp. 배양액으로부터 아가레이즈 분리를 위한 음이온교환 크로마토그래피 최적화)

  • Kim, Yu-Na;Lee, Jae-Ran;Kim, Mu-Chan;Kim, Sung-Bae;Chang, Yong-Keun;Hong, Soon-Kwang;Kim, Chang-Joon
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.840-845
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    • 2011
  • Degradation products of agarose are biologically active and thus used as an ingredient in pharmaceuticals or functional cosmetics. Furthermore, it has been strongly considered as a substrate for bio-ethanol fermentation. Recently, we isolated new agarase-producing microorganism, Pseudoalteromonas sp. from south sea of Korea. In this study, we aimed to separate and purify the agarase from culture broth of this strain. Separation of agarase was performed by ion- exchange chromatography on DEAE-Sepharose resin. Equilibrium pH and volume ratio of resin to the amount of protein were optimized for the efficient adsorption of protein. 410 ${\mu}g$ of protein was completely adsorbed to 3 mL of resin at pH 7.5. The total amount of eluted protein increased as NaCl concentration increased to 400 mM at isocratic elution. Agarase was separated by linear gradient elution of NaCl (0~1,000 mM). Three major protein peaks were observed and the presence or absence of agarase in these eluted proteins was measured by Lugol's staining technique. Only six eluted protein fractions showed strong agarase activity.

Ginsenoside Rh2 attenuates microglial activation against toxoplasmic encephalitis via TLR4/NF-κB signaling pathway

  • Xu, Xiang;Jin, Lan;Jiang, Tong;Lu, Ying;Aosai, Fumie;Piao, Hu-Nan;Xu, Guang-Hua;Jin, Cheng-Hua;Jin, Xue-Jun;Ma, Juan;Piao, Lian-Xun
    • Journal of Ginseng Research
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    • v.44 no.5
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    • pp.704-716
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    • 2020
  • Background: Ginsenoside Rh2 (GRh2) is a characterized component in red ginseng widely used in Korea and China. GRh2 exhibits a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer properties. However, its effects on Toxoplasma gondii (T. gondii) infection have not been clarified yet. Methods: The effect of GRh2 against T. gondii was assessed under in vitro and in vivo experiments. The BV2 cells were infected with tachyzoites of T. gondii RH strain, and the effects of GRh2 were evaluated by MTT assay, morphological observations, immunofluorescence staining, a trypan blue exclusion assay, reverse transcription PCR, and Western blot analyses. The in vivo experiment was conducted with BALB/c mice inoculated with lethal amounts of tachyzoites with or without GRh2 treatment. Results and conclusion: The GRh2 treatment significantly inhibited the proliferation of T. gondii under in vitro and in vivo studies. Furthermore, GRh2 blocked the activation of microglia and specifically decreased the release of inflammatory mediators in response to T. gondii infection through TLR4/NF-κB signaling pathway. In mice, GRh2 conferred modest protection from a lethal dose of T. gondii. After the treatment, the proliferation of tachyzoites in the peritoneal cavity of infected mice markedly decreased. Moreover, GRh2 also significantly decreased the T. gondii burden in mouse brain tissues. These findings indicate that GRh2 exhibits an antieT. gondii effect and inhibits the microglial activation through TLR4/NF-κB signaling pathway, providing the basic pharmacological basis for the development of new drugs to treat toxoplasmic encephalitis.

Preparation of Hydrophobic Antimicrobal Compounds Encapsulated Nanoparticles Using Alkoxysilane-functionalized Amphiphilic Polymer Precursor and Their Antimicrobial Properties (실란 기능화 양친성 고분자 전구체를 이용한 소수성 항균물질 담지 나노 입자 제조 및 항균 특성)

  • Kim, Nahae;Kim, Juyoung
    • Journal of Adhesion and Interface
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    • v.18 no.1
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    • pp.13-24
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    • 2017
  • In this study, nanoparticles which encapsulated hydrophobic antimicrobial compounds with 50wt% of payload and 70%of solid content were prepared. These nanoparticles could be dispersed at water as well as various medium. Water dispersible organic-inorganic (O-I) hybrid nanoparticles were first prepared using alkoxysilane-functionalized amphiphilic polymer precursors through a conventional sol-gel process. Hydrophobic antimicrobial compound, Eugenol encapsulated nanoparticles were prepared using these O-I hybrid nanoparticles through a new nanoprecipitation process. The effect of various preparation on the size of nanoparticles, amount of payload, antimicrobial activity, and release rate of encapsulated compounds was investigated. All eugenol-encapsulated O-I nanoparticles regardless of preparation condition showed the same minimal inhibitory concentration (MIC) (50mg/ml) and 99% of antimicrobial activity for every strain. Their antimicrobial activity could maintain longer than two weeks. Especially, eugenol-encapsulated O-I nanoparticles prepared using tetraethoxysilane (TEOS) exhibited the highest payload (50wt%) and the lowest release rate which was owing to higher inorganic content in the O-I nanoparticles. And these O-I nanoparticles dispersed in hexanediol (HD) showed the highest antimicrobial activity and solid content (70wt%) because HD acted as a solvent as well as a antimicrobial agent.