• Title/Summary/Keyword: isolation techniques

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Antioxidant, antinociceptive activity and general toxicity study of Dendrophthoe falcata and isolation of quercitrin as the major component

  • Hasan, Md Shihab;Ahmed, Md Iqbal;Mondal, Sukla;Masud, Mohammad Methedi;Sadhu, Samir Kumar;Ishibashi, Masami;Uddin, Shaikh Jamal
    • Advances in Traditional Medicine
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    • v.6 no.4
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    • pp.355-360
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    • 2006
  • Plants have been used in traditional medicinal system for centuries. Bangladeshi medicinal plants have received considerable attention from the researchers for evaluation of their bioactivity. As a part of our ongoing research of screening the Bangladeshi medicinal plants, the ethanolic extract of Dendrophthoe falcata have been chosen for the present study. The ethanolic extract of the leaves of the plant have been assessed for their antioxidant, antinociceptive, and general toxicity. The extract showed potent antioxidant activity ($IC_{50}5.1{\mu}g/ml$) using DPPH radical scavenging assay, which is comparable to the standard ascorbic acid ($IC_{50}4.6{\mu}g/ml$). The extract significantly and dose dependently inhibited the acetic acid induced writhing in mice (71.2%, P < 0.001 and 28.0%, P < 0.05 for 500 and 250 mg/kg body weight, respectively). A general toxicity was assessed by a simple and low cost assay using brine shrimp lethality as an indicator. The extract showed low level of toxicity ($LC_{50}100{\mu}g/ml$). Using different chromatographic techniques, quercitrin (quercetin 3-O-${\alpha}$-rhamnoside) was separated as the major component from the extract. The structure was elucidated by detailed 1D and 2D NMR and mass spectral analysis.

Isolation and Structure Identification of Phytotoxins from Lasiodiplodia theobrorme, the Cause of JAVA Black Rot of Sweet Potato (고구마 검은썩음병균(Lasiodiplodia theobromae)으로부터 식물독소의 분리 및 구조)

  • Lim, Chi-Hwan
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.118-123
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    • 1998
  • Lasiodiplodia theobrorme is a pathogen of Java black rot on sweet potato. This fungus infects the tuberous root during storage after harvest. Invasion of the fungus results in the expansion of necrosis sites into the tuberous roots. The resultant necrotic symptom of the tissue is also induced by application of acetone extract of the fungus growing on potato sucrose agar (PSA) culture. The active principles to induce the necrosis are purified from the acetone extract as follows. After evaporation of hexane-benzene-EtOAc (1:2:1, v/v/v) the extract was fractioned on silica gel column, using a solvent gradient system from n-hexane to EtOAc and then to MeOH. The active fractions were purified with HPLC on Nucleosil 50-5 column by eluting n-hexane to EtOAc. Their structures are established by using spectroscopic techniques and synthesis to 4-hydroxymellein and mellein, respectively. Application of small amount of these compounds induce expansion of the necrotic symptom into the tissue and accumulated ipomeamarone. Conclusively, these compounds acted as phytotoxins (inducing necrosis) and as elicitors (eliciting the phytoalexin).

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Isolation of Compounds with Antioxidative Activity from Quickly Fermented Soy-Based Foods

  • Jang, Mi-Young;Cho, Jeong-Yong;Cho, Jeong-Il;Moon, Jae-Hak;Park, Keun-Hyung
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.214-219
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    • 2006
  • A bacterial strain, initially identified as B1-3, was isolated from cheonggukjang, a traditional Korean dish made from fermented soybeans. Using the Biolog system and 16S rRNA sequence analysis, we identified B1-3 as Bacillus mojavensis. We manufactured a quickly fermented soybean (QFS) food product using the B. mojavensis, and guided by their 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging ability. We isolated substances with antioxidative activity from it. Using mass spectrometry (MS) and nuclear magnetic resonance (NMR) techniques, we isolated 4 compounds from the ethyl acetate (EtOAc)-soluble neutral fraction of methyl alcohol (MeOH) extracts of the QFS food product (genistein, daidzein, 3R,4R-3-methyl-3,4-dihydroxy-2-pentanone, and 3S,4R-3-methyl-3,4-dihydroxy-2-pentanone) and 3 compounds from its acidic fraction (4-hydroxyphenylacetic acid, genistin, and daidzein). Two compounds from the neutral fraction (3R,4R-3-methyl-3,4-dihydroxy-2-pentanone and 3S,4R-3-methyl-3,4-dihydroxy-2-pentanone) were not detected in nonfermented soybeans (NFS) or in the filtrate of the LB broth used to culture B. mojavensis. However, they were detected in the filtrate of the same broth when it contained 2% glucose. These results suggest that these 2 compounds were derived from glucose (or other saccharides) in the soybean during fermentation. One compound that was found in the acidic fraction (4-hydroxyphenylacetic acid) was readily detected in NFS, but not in the culture broth. This suggests that 4-hydroxyphenylacetic acid was derived from NFS. We concluded that the antioxidative activity of cheonggukjang is a result of the interactions between soybean components and the microorganisms used in the fermentation of cheonggukjang.

Analysis of Ecological Variation after Creation of the Eco-pond (생태연못 조성공법 적용후의 자연생태 변화분석)

  • Lee, Eun Yeob;Moon, Seok Ki
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.4 no.1
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    • pp.1-15
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    • 2001
  • The purpose of this study is to evaluate the creation techniques of eco-pond, one of biotopes to promote biodiversity in urban residence area. Investigation were classified out plant, mammals, amphibia, reptiles, birds, fishes and insects. The results were summarized as follows: Around the eco-pond shows simple vegetation structurs, consisted of Pinus densoflora S et Z. and Robinia pseudoacacia under competition. In case of shrub, consisted of 4 species but plant growing appearence diversely by seasons. The evaluation of vegetation of eco-pond, there are found 4 species of aquatic plants. Inside the revetment of pond, Echinochloa crus-galli, Persicaria hydropiper, Digiaria sanguinalis, Cyperus microiria and Bidens frondosa L. are mainly distributed. Near the revetment, Trifolium repens L. and Digiaria sanguinalis are prevailed. And in its background, Erigeron canadensis, Erigeron annuus and vines are begins to make their appearances. When evaluation animals in eco-pond and contrast plot, it show simple species and numbers of mammals. It seemed to be resulted from its isolation and outside intervention by users In eco-pond, Pica pica and Streptopelia orientalis are mainly found and in contrast plot of Columba livia, which are so strong adaptation to city life environment. In case of amphibia and reptiles, none is observed in contrast plot, but in ecological pond, Rana nigromaculata and Hyla japonica are constantly observed. In case of insects, more species are found in eco-pond than contrast plot. And in eco-pond, more dragonflies are visibly increased one year after its construction. In floral zone inside of pond revetment, grasshopper and Locusta migratoria are frequently observed. In case of butterflies, they are mainly found in log fence and willow(salix) around eco-pond. In case of fishes inside of eco-pond, the species and its density are remarkable increased one year after the construction. With above evaluation results, we have identify the increase effect of biodiversity after construction of the eco-pond.

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Application of Embryo Transfer Technology (수정란 이식 기술의 응용)

  • Lim, Hyun-Joo;Son, Jun-Kyu;Yoon, Ho-Beak;Baek, Kwang-Soo;Choe, Chang-Yong;Kim, Sidong;Kwon, Eung-Gi
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.163-168
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    • 2013
  • Embryo transfer (ET) technology is of high importance in modern cattle breeding programs. ET is one step in the process of removing one or more embryos from the reproductive tract of an outstanding donor female and transferring them to one or more recipient females. Embryos also can be produced in the laboratory via techniques such as in vitro fertilization (IVF). But the actual transfer of an embryo is only one step in a series of processes that may include some or all of the following: superovulation and insemination of donors, collection of embryos, isolation, evaluation and short-term storage of embryos, micromanipulation and genetic testing of embryos, freezing of embryos and embryo transfer. Cryopreservation and direct transfer of frozen-thawed embryos is common-place with pregnancy rates near that of fresh embryos. Polymerase chain reaction (PCR) technology is currently being used for sexing embryos, and this technology will be used for "embryo diagnostics" and "embryo genomics" in the future. Although, many limitations and problems remain to overcome, these and other new technologies promise to change livestock breeding drastically in the next decade.

Discovery of antigens for early detection of Mycobacterium avium subsp. paratuberculosis and analysis of characteristics using bioinformatics tools (Mycobacterium avium subsp. paratuberculosis 감염 초기 개체 검출을 위한 항원 탐색 및 특성 분석)

  • Park, Hong-Tae;Park, Hyun-Eui;Shin, Min-Kyoung;Cho, Yong-Il;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.89-95
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    • 2015
  • Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is one of the most widespread and economically important diseases in cattle. Current diagnostic methods are based on the detection of anti-MAP antibodies in serum or isolation of the causative agent. However, these techniques are often not applicable for cases of subclinical infection due to relatively low sensitivity. Therefore, finding new antigen candidates that strongly react with the host immune system had been attempted. To effectively detect infection during the subclinical stage, several antigen candidates were selected based on previous researches. Characteristics of the selected antigen candidates were analyzed using bioinformatics-based prediction tools. A total of nine antigens were selected (MAP0862, MAP3817c, MAP2077c, MAP0860c, MAP3954, MAP3155c, MAP1204, MAP1087, and MAP2963c) that have MAP-specific and/or high immune responses to infected animals. Using a transmembrane prediction tool, five of the nine antigen candidates were predicted to be membrane protein (MAP3817c, MAP3954, MAP3155c, MAP1087, and MAP1204). Some of the predicted protein structures identified using the I-TASSER server shared similarities with known proteins found in the Protein Data Bank database (MAP0862, MAP1204, and MAP2077c). In future studies, the characteristics and diagnostic efficiency of the selected antigen candidates will be evaluated.

Enhancing e-Learning Interactivity vla Emotion Recognition Computing Technology (감성 인식 컴퓨팅 기술을 적용한 이러닝 상호작용 기술 연구)

  • Park, Jung-Hyun;Kim, InOk;Jung, SangMok;Song, Ki-Sang;Kim, JongBaek
    • The Journal of Korean Association of Computer Education
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    • v.11 no.2
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    • pp.89-98
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    • 2008
  • Providing appropriate interactions between learner and e- Learning system is an essential factor of a successful e-Learning system. Although many interaction functions are employed in multimedia Web-based Instruction content, learner experience a lack of similar feedbacks from educators in real- time due to the limitation of Human-Computer Interaction techniques. In this paper, an emotion recognition system via learner facial expressions has been developed and applied to a tutoring system. As human educators do, the system observes learners' emotions from facial expressions and provides any or all pertinent feedback. And various feedbacks can bring to motivations and get rid of isolation from e-Learning environments by oneself. The test results showed that this system may provide significant improvement in terms of interesting and educational achievement.

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Quinone Reductase Induction Activity of Phlorotannins Derived from Eisenia bicyclis in Hepa1c1c7 Cells

  • Yoon, Na Young;Lee, Sang-Hoon;Shim, Kil Bo;Lim, Chi-Won;Lee, Moon-Hee;Cho, Hyun-Ah;Xie, Chengliang
    • Fisheries and Aquatic Sciences
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    • v.16 no.1
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    • pp.1-5
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    • 2013
  • To assess the feasibility of phlorotannins from Eisenia bicyclis as cancer chemopreventative agents, we tested whether they induced quinone reductase (QR) in Hepa1c1c7 cells. The ethyl acetate (EtOAc) soluble fraction obtained from E. bicyclis exhibited a QR induction activity in Hepa1c1c7 cells. Successive column chromatography of the active EtOAc fraction resulted in the isolation of four phlorotannins. Their structures were elucidated using one- and two-dimensional nuclear magnetic resonance spectroscopic techniques and characterized as phloroglucinol (1), dioxinodehydroeckol (2), dieckol (3), and fucofuroeckol-A (4). Among these compounds, fucofuroeckol-A (4) showed moderate QR induction activity, and dioxinodehydroeckol (2) exhibited potent QR induction potency with $2.05{\pm}0.04$ fold induction at a concentration of $50{\mu}M$ compared to the dimethyl sulfoxide solvent-treated control cells. However, phloroglucinol (1) and dieckol (3) exerted no detectable QR induction activity in Hepa1c1c7 cells. These results suggest that dioxinodehydroeckol could serve as a useful cancer chemopreventive chemical.

Isolation and Characterization of MT2617-2B, a Phospholipase C Inhibitor Produced by an Actinomycetes Isolate (방선균 분리주가 생산하는 Phospholipase C 저해물질인 MT-2617-2B의 분리 및 특성)

  • Ko, Hack-Ryong;Lee, Hyun-Sun;Oh, Won-Keun;Ahn, Soon-Cheol;Kim, Bo-Yeon;Kang, Dae-Ook;Mheen, Tae-Ick;Ahn, Jong-Seog
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.19-26
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    • 1996
  • A phospholipase C (PLC) inhibitor (MT267-2B) was isolated from the culture broth of actinomycetes isolate MT2617-2 by the extraction with n-butanol and column chromatographic techniques. The molecular weight of the inhibitor was 1057, by the spectroscopic analyses of IR, $^{13}C$-and $^{1}H$-NMR and ESI-MS. The chemical structure of MT2617-2B was found to be a macrolide compound consisted of a hemiketal ring, polyhydroxyl and polymethyl groups, which had a malonate and guanidine group as its side chain. MT2617-2B produced its two isomers having the same molecular weight by standing in methanol solution at room temperature. Therefore, MT2617-2B was identified as copiamycin and niphithricin A, macrolide antibiotics. The values of $IC_{50}$ against PLC-${\gamma}$1 and PLC-${\beta}$1 were 25 and 50${\mu}$g/ml, respectively. MT2617-2B had antimicrobial activities against Staphylococcus aureus and Candida albicans, but not against Escherichia coli.

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Candida non albicans with a High Amphotericin B Resistance Pattern Causing Candidemia among Cancer Patients

  • Kalantar, Enayatollah;Assadi, Mojan;Pormazaheri, Helen;Hatami, Shiva;Barari, Maryam Agha;Asgari, Esfandiar;Mahmoudi, Elaheh;Kabir, Kourosh;Marashi, Seyed Mahmoud Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10933-10935
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    • 2015
  • Background: Many scientists have reported Candida species to be of great concern because of the high frequency that they colonize and infect human hosts, particularly cancer patients. Moreover, in the last decades Candida species have developed resistance to many antifungal agents. Based on this, we aimed to identify and determine the prevalence of Candida spp from blood culture bottles among cancer patients and their antifungal resistance pattern. Materials and Methods: From the blood culture bottles isolation and identification of the Candida spp were performed by conventional microbiological techniques. The in vitro antibiotic resistance pattern of the isolates was determined by CLSI guidelines. Genomic DNA was isolated and amplified. Each gene was separated by agar gel electrophoresis. Results: Identification of Candida spp was based on the presence of yeast cells in direct examination, culture and DNA extraction. Of the 68 blood samples collected during the study period (April 2013 to October 2013), five (7.35%) were positive for the presence of Candida spp, 2 (40%) of which were identified as Candida albicans and 3 (60%) were Candida non-albicans. Conclusions: High resistance to amphotricin B was observed among all the Candida non-albicans isolates. Regular investigations into antifungal resistance will help us to get an updated knowledge about their antibiotic resistance pattern which may help the physician in selecting the antibiotics for empirical therapy.