• Title/Summary/Keyword: Liquid Medium

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Neuroprotective effects of phenolic compounds isolated from Spiraea prunifolia var. simpliciflora (조팝나무(Spiraea prunifolia var. simpliciflora)로부터 분리한 페놀 화합물의 신경세포 보호효과)

  • Oh, Seon Min;Choi, Doo Jin;Kim, Hyoung-Geun;Lee, Jae Won;Lee, Young-Seob;Lee, Jeong-Hoon;Lee, Seung-Eun;Kim, Geum-Soog;Baek, Nam-In;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.61 no.4
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    • pp.397-403
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    • 2018
  • The leaves of Spiraea prunifolia were extracted with 80% aqueous MeOH and the concentrates were partitioned into EtOAc, n-BuOH, and $H_2O$ fractions. The repeated $SiO_2$ or ODS column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of two phenolic glucosides. The chemical structures of these compounds were determined as isosalicin (1) and crenatin (2) based on spectroscopic analyses including Nuclear magnetic resonance and MS. Extracts were analyzed using UPLC-MS/MS providing a short analysis time within 5 min using MRM technique. The concentration of crenatin was higher as 9.53 mg/g and isosalicin was lower as 0.65 mg/g. Neuroprotective effects of these compounds against hydrogen peroxide ($H_2O_2$)-induced neurotoxicity were evaluated. The results showed that exposure to $H_2O_2$ induced morphological changes, cell death and neurotoxicity in SK-N-MC cells. However, pretreatment with crenatin resulted in inhibition of morphological change, reduction of loss of cell viability and attenuation of neuronal damage. These results suggested that neuroprotective effect of crenatin isolated from S. prunifolia can be a good candidate for the development of health beneficial foods which can ameliorate the degenerative neuronal disease caused by oxidative stress.

Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3 (해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특성)

  • Kim, Da Som;Kim, Jong-Hee;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.329-336
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    • 2021
  • Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 ℃. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

Effect of Anti-inflammation on Oryeong-san Formulation for Mix Extract Tablet (오령산 정제 개발 및 항염증 효과)

  • Kim, Se Jin;Leem, Hyun Hee;Nam, Won Hee;Son, Su Mi;Choi, Hye Min;Kim, Myung Jin;Kim, Jung Ok;Lee, Hwa Dong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.34 no.6
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    • pp.348-354
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    • 2020
  • Oryeong-san (ORS) is a traditional Korean herbal medicine widely used for renal associated diseases, composed of five medicine herbs; Atractylodes japonica Koidzumi, Cinnamomum cassia Presl, Polyporus umbellatus Fries, Poria cocos Wolf and Alisma orientale Juzepzuk. We studied to improve the convenience of intake and portability by developing modernized dosage forms, and examined the effect on anti-inflammation of ORS. In order to develop the tablet formulation of ORS (ORS-F), the tablets were evaluated on the basis of physical characteristics include diameter, thickness, weight variation, hardness, friability and disintegration. To analyze the marker components of ORS-F, eight index markers from five herbal medicines were chosen. And the method using high performance liquid chromatography (HPLC) with diode-array detector method was established for the simultaneous analysis. The biological activities were examined the effect of ORS-F on pro-inflammation mediated by LPS-stimulation. The production of nitric oxide (NO) and cytokines were determined by reacting cultured medium with griess reagent and enzyme-linked immunosorbent assay (ELISA). The expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) were investigated by Western blot and RT-PCR. The anti-oxidant activities of OJS-F increased markedly, in a dose-dependent manner. and, The total phenolic compound and flavonoids contents of OJS-F were 10.20±0.09 ㎍/㎎ and 12.86±0.86 ㎍/㎎. OJS-F which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) from the RAW264.7 macrophages. Therefore, the developed formulation for tablet of ORS-F provide efficiency and usability, and indicated effect of anti-inflammation.

A simple mid-term preservation method (SMPM) of plant callus under low temperature conditions (저온 보존을 이용한 간편 중기 식물캘러스 저장법)

  • Park, Sung-Chul;Park, Su Hyun;Kim, Soyoung;Jeong, Yu Jeong;Kim, Cha Young;Jeong, Jae Cheol
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.187-192
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    • 2022
  • The repeated monthly or weekly subculture of plant callus is labor intensive and increases the risk of somaclonal variation from the parental callus line. The most effective method for preserving plant callus is cryopreservation, which involves storage in liquid nitrogen. However, this method cannot be applied to the callus of different plant species in the same manner, so it is difficult to develop a standardized cryopreservation method. In addition, the survival rate of the frozen callus after thawing and the regeneration rate after survival are uncertain. Therefore, it is necessary to develop a method to extend the subculture interval of plant callus in an active state. In this study, active plant calli of various species without freezing was incubated at 15℃ for 4 to 12 weeks without subculture. After 12 weeks, 8 lines of plant callus grew less than 2-fold when cultured at 25℃, but at least 2 times as much when cultured at 15℃. Moreover, total antioxidant activity did not differ significantly between plant callus recovered at 25℃ after culturing at 15℃ or at 25℃. These results show that the subculture interval can be extended at a temperature of 15℃ without need for modified medium composition or additional processes. In addition, positive results in all calli of several plant species are expected to reduce labor as well as somaclonal variation by increasing the subculture.

Expression and Localization of Heat Shock Protein 70 in Frozen-Thawed IVF and Nuclear Transfrred Bovine Embryos

  • Park, Y.J;S.J Song;J.T Do;B.S Yoon;Kim, A.J;K.S Chung;Lee, H.T
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.78-78
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    • 2002
  • The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

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Artificial Cultivation and Antioxidant Activity of Korean Native Mushroom Pycnoporus coccineus (한국산 간버섯의 인공재배 및 항산화 활성 평가)

  • Lee, Y.K.;Kim, M.K.;Yun, B.S.;Seo, G.S.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.21 no.2
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    • pp.35-47
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    • 2019
  • The mycelial growth of P. coccineus strain was good in PDA and YMA, but mycelial growth was low in MEA. Light irradiation during the incubation period affected the pigment formation and density of mycelia. Mushroom of P. coccineus strain was able to produce fruiting bodies in both bottle and bag cultivation, and oak sawdust was found to be the most suitable substrate for spawn culture and cultivation. In artificial cultivation using sawdust medium, fruiting body was grown to the extent that visual observation was possible from the 15th day, and it formed about 5 days fast in the treatment group with low relative humidity. From 40 to 45 days of mushroom development, mature fruiting bodies could be harvested, and the lower relative humidity of the growing room favored mushroom development and growth. Antioxidant activity of fruiting bodies harvested from artificial cultivation showed that ABTS radical scavenging activity of bottle-cultivated and wild fruit bodies were shown at 505㎍/㎖ and 515㎍/㎖, respectively. However, fruiting bodies harvested in bag cultivation were high at 910㎍/㎖. As a result of DPPH radical scavenging activity, all extracts were found to be inactive, exhibiting IC50 value of more than 2,000㎍/㎖ concentration. The ethyl acetate extract of mushrooms obtained from bottle cultivation showed the highest activity with 1,550㎍/㎖ IC50 value. Methanol extract of wild fruit bodies had the highest ABTS radical scavenging activity at the same concentration (10mg/㎖).

A Study on the Antifungal Properties of Ranunculaceae Herbal Medicines (미나리아재비과 한약재의 항진균성 검증 및 비교 연구)

  • Jae-yeoup Lim;Sang Ki Choi
    • Journal of Life Science
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    • v.34 no.6
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    • pp.393-398
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    • 2024
  • The purpose of this study was to verify the antifungal properties of various herbal medicines belonging to the Ranunculaceae family and to find an extraction method effective in inhibiting fungal growth. When antifungal activity was measured in a liquid medium with extracts obtained by either hot water extraction or organic solvent extraction of the herbal medicines Clematis apiifolia, Coptis chinensis, and Pusatilla chinensis, effective results were obtained from the chloroform extract. In addition, fungal growth inhibition experiments were performed on unicellular fungi, Candida albicans, Candida tropicalis, and Candida lusitaniae, and on filamentous fungi, such as Pythium ultimum, Aspergillus fumigatus, and Fusarium oxysporum, using disk diffusion experiments on solid media. It was confirmed that P. chinensis extract has excellent antifungal properties against Candida spp. and C. apiifolia extract against filamentous mold. Finally, GC-MS analysis was performed to explore the useful antifungal substances present in the extract. As a result of the study, thurbergenone from C. apiifolia and 16-hydroxycleroda-3, 13(14)-dien-15, 16-olide (16-HCDO) from C. chinensis were confirmed as antifungal candidates. In conclusion, it was confirmed that C. apiifolia, C. chinensis, and P. chinensis have antifungal activity against various fungi, and in GC-MS analysis, all herbal medicines were confirmed to have different antifungal candidates. These results indicate that the Ranunculaceae family has evolved in several directions for fungal resistance traits.

Computational Fluid Dynamics Study of Channel Geometric Effect for Fischer-Tropsch Microchannel Reactor (전산유체역학을 이용한 Fischer-Tropsch 마이크로채널 반응기의 채널 구조 영향 분석)

  • Na, Jonggeol;Jung, Ikhwan;Kshetrimayum, Krishnadash S.;Park, Seongho;Park, Chansaem;Han, Chonghun
    • Korean Chemical Engineering Research
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    • v.52 no.6
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    • pp.826-833
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    • 2014
  • Driven by both environmental and economic reasons, the development of small to medium scale GTL(gas-to-liquid) process for offshore applications and for utilizing other stranded or associated gas has recently been studied increasingly. Microchannel GTL reactors have been prefrered over the conventional GTL reactors for such applications, due to its compactness, and additional advantages of small heat and mass transfer distance desired for high heat transfer performance and reactor conversion. In this work, multi-microchannel reactor was simulated by using commercial CFD code, ANSYS FLUENT, to study the geometric effect of the microchannels on the heat transfer phenomena. A heat generation curve was first calculated by modeling a Fischer-Tropsch reaction in a single-microchannel reactor model using Matlab-ASPEN integration platform. The calculated heat generation curve was implemented to the CFD model. Four design variables based on the microchannel geometry namely coolant channel width, coolant channel height, coolant channel to process channel distance, and coolant channel to coolant channel distance, were selected for calculating three dependent variables namely, heat flux, maximum temperature of coolant channel, and maximum temperature of process channel. The simulation results were visualized to understand the effects of the design variables on the dependent variables. Heat flux and maximum temperature of cooling channel and process channel were found to be increasing when coolant channel width and height were decreased. Coolant channel to process channel distance was found to have no effect on the heat transfer phenomena. Finally, total heat flux was found to be increasing and maximum coolant channel temperature to be decreasing when coolant channel to coolant channel distance was decreased. Using the qualitative trend revealed from the present study, an appropriate process channel and coolant channel geometry along with the distance between the adjacent channels can be recommended for a microchannel reactor that meet a desired reactor performance on heat transfer phenomena and hence reactor conversion of a Fischer-Tropsch microchannel reactor.

Characteristics of Membrane Permeability on the Separation of Solid in a Liquid Livestock Manure (축분액비의 고액분리에 있어서 분리막의 투과특성)

  • 황명구;차기철;이명규
    • Journal of Animal Environmental Science
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    • v.6 no.3
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    • pp.175-184
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    • 2000
  • A lab-scale MF membrane reactor was installed to investigate the membrane permeability, characteristics of membrane fouling at each conditions, and quality of permeate (liquid livestock manure) in the separation of solid-matters using membrane. Experiment was divided three filtration type such as follows; continuous filtration, gravity filtration, and intermittent filtration. As a result of experiment, flux 1 LMH was maintained for 7days, and trans-membrane pressure(TMP) was increased gradually under 10cmHg, but it was increased immediately after 10cmHg, respectively. However, the flux was increased, the Tmax was decreased exponential more and more. During the pure-flux test, most of the fouling of membrane was reversible. At the gravity filtration, permeate could be obtained as 1.75 LMH for 3.5days without any other electronic pressure. As an investigation of membrane surface, this study could be decided that the reason of fouling at the lower flux (Run 1 and 2) was attached matters in membrane surface, but at the higher flux (Run 4-6) was concentration polarization.

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Biosynthesis of the extracellular enzymes in de novo during the differentiation of Aspergillus niger (검정곰팡이의 형태분화에 따른 세포외성효소의 신생적생합성에 관한 연구)

  • Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.6 no.2
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    • pp.1-10
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    • 1978
  • In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.

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