• Title/Summary/Keyword: In vitro screening

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Isolation of MLL1 Inhibitory RNA Aptamers

  • Ul-Haq, Asad;Jin, Ming Li;Jeong, Kwang Won;Kim, Hwan-Mook;Chun, Kwang-Hoon
    • Biomolecules & Therapeutics
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    • v.27 no.2
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    • pp.201-209
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    • 2019
  • Mixed lineage leukemia proteins (MLL) are the key histone lysine methyltransferases that regulate expression of diverse genes. Aberrant activation of MLL promotes leukemia as well as solid tumors in humans, highlighting the urgent need for the development of an MLL inhibitor. We screened and isolated MLL1-binding ssRNAs using SELEX (${\underline{S}}ystemic$ ${\underline{E}}volution$ of ${\underline{L}}igands$ by ${\underline{E}}xponential$ enrichment) technology. When sequences in sub-libraries were obtained using next-generation sequencing (NGS), the most enriched aptamers-APT1 and APT2-represented about 30% and 26% of sub-library populations, respectively. Motif analysis of the top 50 sequences provided a highly conserved sequence: 5'-A[A/C][C/G][G/U][U/A]ACAGAGGG[U/A]GG[A/C] GAGUGGGU-3'. APT1, APT2, and APT5 embracing this motif generated secondary structures with similar topological characteristics. We found that APT1 and APT2 have a good binding activity and the analysis using mutated aptamer variants showed that the site information in the central region was critical for binding. In vitro enzyme activity assay showed that APT1 and APT2 had MLL1 inhibitory activity. Three-dimensional structure prediction of APT1-MLL1 complex indicates multiple weak interactions formed between MLL1 SET domain and APT1. Our study confirmed that NGS-assisted SELEX is an efficient tool for aptamer screening and that aptamers could be useful in diagnosis and treatment of MLL1-mediated diseases.

A Review of Research Trends in Traditional Korean Medicine for Nonalcoholic Fatty Liver Disease (비알코올 지방간질환에 대한 한의학 연구 동향 보고: 국내 논문을 중심으로)

  • Kim, Na-yeon;Yoon, Mi-jung;Choi, Hong-sik;Kim, Seung-mo;Kim, Sang-chan;Kim, Kyung-soon
    • The Journal of Internal Korean Medicine
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    • v.43 no.4
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    • pp.680-719
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    • 2022
  • Objectives: The aim of this study was to investigate the trends in research on non-alcoholic fatty liver disease (NAFLD) using Korean traditional medicine. Methods: This review included studies related to the treatment of NAFLD, irrespective of the year of publication. The search terms were "nonalcoholic fatty liver disease", "non-alcoholic fatty liver", "nonalcoholic hepatitis", "herb", "herbal medicine", "acupuncture", and "traditional medicine". The studies were analyzed according to the type of research. Results: After screening, 179 studies were selected from the 592 identified by the search. The types of studies were 151 in vivo or in vitro studies, 5 randomized controlled trials, 12 case reports, 2 unspecified clinical studies, 8 review articles, and 1 article that was difficult to classify. Conclusion: Analysis of the trends in Korean traditional medicine treatment by reviewing the studies on NAFLD revealed a focus on experimental studies rather than clinical studies. Therefore, multifaceted and well-designed studies are needed.

Antimicrobial and antioxidant activity of some Indian medicinal plants for the protection against fish pathogenic bacteria

  • Harikrishnan, Ramasamy;Jawahar, Sundaram;Kim, Man-Chul;Kim, Ju-Sang;Jang, Ik-Soo;Balasundaram, Chellam;Heo, Moon-Soo
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.317-326
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    • 2009
  • This study has shown the screening of anti-bacterial activity of three Indian medicinal plant choloroform : methanol (50:50) solvent leaf extracts (i.e. Azadirachta indica, Ocimum sanctum, and Curcuma longa) with different concentrations (10, 5, 2.5, 1.25, 0.625, 0.312, and 0.156 mg/ml) under in vitro conditions against fish pathogenic bacteria, Aeromonas hydrophila, Streptococcus iniae, Vibrio harveyi, V. anguillarum, and Edwardsiella tarda isolated from olive flounder farms, Jeju Island, South Korea. The anti-microbial activity of the A. indica and O. sanctum extracts yielded the zones of growth inhibition (ZI) was 3 and 1mm against A. hydrophila at concentration of 0.156 mg/ml when compared to that of tetracycline standard (3 mm). At highest concentration (10 mg/ml) of A. indica, O. sanctum, and C. longa, high inhibition was 9, 7, and 6 mm when compared to that of tetracycline (11 mm) against A. hydrophila. The minimum inhibitory concentration (MIC) of A. indica, O. sanctum, and C. longa at 0.156 mg/ml that yield 9, 10, and 13 CFU/ml for A. hydrophila, 16, 22, and 25 CFU/ml for S. iniae and 18, 22, and 23 CFU/ml for E. tarda compared to the tetracycline. At highest concentration (10 mg/ml) of the three extracts was better inhibiting the growth of A. hydrophila, S. iniae and E. tarda. A. indica, O. sanctum, and C. longa were determined to the potential antioxidant activityon the basis of their scavenging activity of the stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical. A. indica extract was 0.625 mg/ml which indicated that the strong anti-oxidant activity. However, O. sanctum and C. longa extracts showed weak anti-oxidant activity at this concentration. Hence, in vitro assay among the pathogens, A. hydropila is better inhibitory activity of the extracts. It is evident that the Indian medicinal plants extracts were subjected to its effectiveness against A. hydrophila, S. iniae, and E.tarda at low concentrations. The obtained results in the present study suggested that the Indian plant extracts is a prevention tools for Korean olive flounder aquaculture pathogens and its need further advance investigation.

NgR1 Expressed in P19 Embryonal Carcinoma Cells Differentiated by Retinoic Acid Can Activate STAT3

  • Lee, Su In;Yun, Jieun;Baek, Ji-Young;Jeong, Yun-Ji;Kim, Jin-Ah;Kang, Jong Soon;Park, Sun Hong;Kim, Sang Kyum;Park, Song-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.2
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    • pp.105-109
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    • 2015
  • NgR1, a Nogo receptor, is involved in inhibition of neurite outgrowth and axonal regeneration and regulation of synaptic plasticity. P19 embryonal carcinoma cells were induced to differentiate into neuron-like cells using all trans-retinoic acid and the presence and/or function of cellular molecules, such as NgR1, NMDA receptors and STAT3, were examined. Neuronally differentiated P19 cells expressed the mRNA and protein of NgR1, which could stimulate the phosphorylation of STAT3 when activated by Nogo-P4 peptide, an active segment of Nogo-66. During the whole period of differentiation, mRNAs of all of the NMDA receptor subtypes tested (NR1, NR2A-2D) were consistently expressed, which meant that neuronally differentiated P19 cells maintained some characteristics of neurons, especially central nervous system neurons. Our results suggests that neuronally differentiated P19 cells expressing NgR1 may be an efficient and convenient in vitro model for studying the molecular mechanism of cellular events that involve NgR1 and its binding partners, and for screening compounds that activate or inhibit NgR1.

Effect of various chromatographic terpenoid fractions of Luffa cylindrica seeds on in-vitro antimicrobial studies

  • Nagarajan, K.;Saxena, Pallavi;Mazumder, Avijit;Ghosh, L.K.;Devi, G. Uma
    • Advances in Traditional Medicine
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    • v.10 no.1
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    • pp.21-28
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    • 2010
  • The objective of the present investigation is to evaluate the antimicrobial potency of the terpenoid fractions isolated from Luffa cylindrica seeds against various pathogenic microbes. The seeds were powdered and extracted with methanol in soxhlet appratus based on phytochemical screening. Three terpenoid components were isolated by column chromatography and identified by thin layer chromatography and chemical analysis which were designated as ${LCSF_4}^*$, ${LCSF_6}^*$ & ${LCSF_8}^*$ respectively. Disc diffusion method was employed to determine the antimicrobial effectiveness of test compounds I, II and III $({LCSF_4}^*,\;{LCSF_6}^*\;&\;{LCSF_8}^*)$ against 6 microbial species viz., Staphylococcus (S.) aureus, Bacillus (B.) subtilis, Escherichia (E.) coli, Pseudomonas (P.) aeruginosa, Candida (C.) albicans and Aspergillus niger. The disc was saturated with $100{\mu}l$ of each compound, allowed to dry and introduced on the upper layer of seeded agar plate. The plates were incubated overnight at $37^{\circ}C$. Microbial growth was determined by measuring the zonal inhibition diameters. Compound I showed maximum potency against gram positive S. aureus (21 mm) in comparison with standard ciprofloxacin (38 mm), whereas the same compound was completely devoid of activity against both the fungi tested. Compound II was found to be highly sensitive against both the gram negative E. coli (20 mm) and P. aeruginosa (22 mm). Compound II was found to exhibit maximum potency against the fungi C. albicans (15 mm) and A. niger (20 mm). Compound III was found to be very effective against both the gram positive S. aureus (20 mm) and B. subtilis (15 mm) respectively.

Isolation and Characterization of an Acyclic Isoprenoid from Semecarpus anacardium Linn. and its Antibacterial Potential in vitro - Antimicrobial Activity of Semecarpus anacardium Linn. Seeds -

  • Purushothaman, Ayyakkannu;Meenatchi, Packirisamy;Saravanan, Nallappan;Karuppaiah, Muthu;Sundaram, Ramalingam
    • Journal of Pharmacopuncture
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    • v.20 no.2
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    • pp.119-126
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    • 2017
  • Objectives: Semecarpus anacardium Linn. is a plant well-known for its antimicrobial, antidiabetic and anti-arthritic properties in the Ayurvedic and Siddha system of medicine. This has prompted the screening of this plant for antibacterial activity. The main aims of this study were to isolate compounds from the plant's seeds and to evaluate their antibacterial effects on clinical bacterial test strains. Methods: The n-butanolic concentrate of the seed extract was subjected to thin layer chromatography (TLC) and repeated silica gel column chromatography followed by elution with various solvents. The compound was identified based on observed spectral (IR, $^1H$ NMR, $^{13}C$ NMR and high-resolution mass spectrometry) data. The well diffusion method was employed to evaluate the antibacterial activities of the isolated acyclic isoprenoid compound (final concentration: $5-15{\mu}g/mL$) on four test bacterial strains, namely, Staphylococcus aureus (MTCC 96), Bacillus cereus (MTCC 430), Escherichia coli (MTCC 1689) and Acinetobacter baumannii (MTCC 9829). Results: Extensive spectroscopic studies showed the structure of the isolated compound to be an acyclic isoprenoid ($C_{21}H_{32}O$). Moreover, the isoprenoid showed a remarkable inhibition of bacterial growth at a concentration of $15{\mu}g/mL$ compared to the two other doses tested (5 and $10{\mu}g/mL$) and to tetracycline, a commercially available antibiotic that was used as a reference drug. Conclusion: The isolation of an antimicrobial compound from Semecarpus anacardium seeds validates the use of this plant in the treatment of infections. The isolated compound found to be active in this study could be useful for the development of new antimicrobial drugs.

Screening of Natural Products that have Activities against Skin-Aging (피부노화 억제활성을 갖는 천연 추출물의 탐색)

  • 신지영
    • The Korean Journal of Food And Nutrition
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    • v.14 no.6
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    • pp.568-572
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    • 2001
  • To screen antioxidative and whitening effects of some medicinal plants, 70% ethanol extracts were tested for their electron donating abilities to DPPH ( ${\alpha}$, ${\alpha}$-diphenyl- ${\beta}$-picrylhydrazyl) and inhibitory activities against lipid peroxidation, and tyrosinase inhibition in vitro. The electron donating activities of 70% ethanol extracts at the concentration of 100 $\mu\textrm{g}$/ml were in the order of Forsythiae fructus > Polyporaceae > Glycyrrhizae radix > Schizandrae fructus > Ramulus mori > Carthami semen. The order was Forsythiae fructus > Carthami semen > Schizandrae fructus > Polyporaceae > Glycyrrhizae radix when tested at 10 $\mu\textrm{g}$/ml, indicating that the activities depend on the concentration. The electron donating activities of Forsythiae fructus and Carthami semen at the concentration of 1 $\mu\textrm{g}$/ml were 37% and 31% respectively. Lipid peroxidation inhibitory activities were in the order of Artemisia messerschimidtiana > Glycyrrhizae uadix > Rehmaniae radix > Ramulus mori. Tyrosinase inhibitory activities were in the order of Ramulus mori > Artemisia messerschimidtiana > Oyster mushroom. The results suggest that Forsythiae fructus, Artemisia messerschimidtiana and Ramulus mori could be used as materials for anti-skin aging functional cosmetics.

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Screening of Inhibitory Effect on Aldose Reductase of Vietnam Herbal Medicines (II) (베트남 약용식물 추출물의 알도즈 환원 효소 억제 효능 검색(II))

  • Lee, Yun-Mi;Kim, Jong-Min;Kim, Young-Sook;Jang, Dae-Sik;Kim, Joo-Hwan;Bae, Ki-Hwan;Kim, Jin-Sook
    • Korean Journal of Pharmacognosy
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    • v.39 no.4
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    • pp.324-329
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    • 2008
  • Aldose reductase (AR), the key enzyme in the polyol pathway, plays an important role in the pathogenesis of diabetic complications such as cataract formation. AR inhibitors (ARI) can prevent or reverse early abnormalities in diabetic complications. However, none of ARI has achieved worldwide use because of limited efficacy or undesirable side effects. Therefore, evaluating natural sources for ARI potential may lead to the development of safer and more effective agents against diabetic complications. 49 Vietnam herbal medicines have been investigated with an in vitro evaluation system using aldose reductase inhibitory activities. Of these, 7 herbal medicines, Artemisia annua (leaf, stem), Duranta repens (leaf, stem), Excoecaria cochinchinensis (aerial part), Flueggea virosa (leaf, stem), Khaya senegalensis (leaf), Polygonum orientale (aerial part), Toxicodendron succedanea (leaf, stem) exhibited a significant inhibitory activity against aldose reductase. Particularly, Duranta repens (leaf, stem) and Toxicodendron succedanea (leaf, stem) showed two times more potent inhibitory activity than the positive control, 3.3-tetramethyleneglutaric acid (TMG).

Screening for Antioxidant Activity of Edible Plants (식용식물의 항산화효과 검색)

  • 조세연;한용봉;신국현
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.133-137
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    • 2001
  • The antioxidant activitiesof methanol extracts of twenty one vegetables were tested using the method of 1,1-diphenyl-2-picryl hydrazyl (DPPH) reactivity and TBARS substance assay in vitro. The methanol extracts of the leaves from three plants such as Aster scaber, Chrysanthemum coronarium, Petasites japonicus were found to be th most effective on DPPH radical scavenging activity. The next effective ones were the leaves of Capsicum annuum, the peels of Ipomoea batatas and Solanum tuberosum, the stems and leaves of Ipomoea batatas, the levels of Solanum melongena, the fruits of Solanum melongena, the green fruits (seed) of Capsicum ammuum in order, and the others did not show a considerable activity. When tested on lipid peroxidation using a lipid peroxidation generation system mediated by $H_{2}O_{2}/Fe^{2+}$ in rat liver homogenates. the peels of Cucumis melo, the leaves and the fruits of Solanum melongena, of Solanum melongena, The levels of Aster scaber and Chrysanthemum coronarium, the stems and leaves of Ipomoea batatas and the peels of Ipomoea batatas were found to exhibit a good protective activity against lipid peroxidatian in the order of effectiveness.

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Screening and Identification of Antifungal Bacillus sp. #72 against the Pathogenic Stem-end Rot of Kiwi Fruit (참다래 꼭지썩음병을 일으키는 Diaporthe actinidiae을 억제하는 길항성 Bacillus sp. #72의 분리 및 동정)

  • Cho, Jung-Il;Cho, Ja-Yong;Park, Yong-Seo;Yang, Seung-Yul;Heo, Buk-Gu
    • The Korean Journal of Community Living Science
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    • v.18 no.2
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    • pp.241-246
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    • 2007
  • This study was conducted to select and screen for an antifungal bacterial strain showing pathogen inhibitory activity against Diaporthe actinidiae, which causes stem-end rot in kiwi fruit. Four bacterial strains were isolated which strongly inhibit Diaporthe actinidiae from among two hundred and fifty bacterial strains screened from the soil where kiwi fruit were grown. By co-culturing bacterial strain #72 and the pathogen causing the stem-end rot of kiwi fruit, bacterial strain #72 showed 81.0% antifungal activity against Diaporthe actinidiae. Bacterial strain #72 was identified to be from the genus Bacillus sp. based on morphological and biochemical characterization. The bacterialization of culture broth for Bacillus sp. #72 which was sterilized at $121^{\circ}C$ for 15 minutes and than purified by $0.45{\mu}m$ membrane filter showed almost all of the antagonistic activity against Diaporthe actinidiae. We have also confirmed that in vitro treatment of Bacillus sp. #72 cultured in SD+B+P(sugar 5%, soy sauce 3%, beef extract 0.2%, peptone 0.2%) medium efficiently inhibited the growth of Diaporthe actinidiae responsible for stem-end rot in kiwi fruit.

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