• Title/Summary/Keyword: liquid cell

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Efficient assay for respiration inhibitor using Saccharomyces cerevisiae (Saccharomyces cerevisiae를 이용한 효율적인 호흡저해제 검정법)

  • Choi, Gyung-Ja;Kim, Jin-Cheol;Kim, Heung-Tae;Cho, Kwang-Yun
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.52-59
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    • 2000
  • A rapid assay to determine respiration inhibition of Saccharomyces cerevisiae by chemicals was developed. S. cerevisiae was harvested with two different liquid media, yeast extract-peptone-dextrose (YPD) medium capable of occurring both glucose fermentation and mitochondrial respiration, and non-fermentable carbon-yeast extract (NFY) medium capable of occurring respiration only Wells in 96-well plate were loaded with each cell suspension and various concentrations of 46 fungicides with various modes of action. n NFY medium, the non-fermentable carbon source, ethanol (NFY-E medium), glycerol (NFY-G medium) or lactate (NFY-L medium), was used. After incubation for $1{\sim}3$ days, minimum inhibitory concentrations (MICs) of the chemicals were recorded in the media. Of the 46 inhibitors employed in this study, four inhibitors of fungal respiration by blockage of electron flux in the mitochondrial respiratory chain, azoxystrobin, kresoxim-methyl, metominostrobin, and trifloxystrobin, exhibited strong antifungal activity in all of NFY media, but no activity in YPD medium. In contrast to this, five N-trihalomethylthio fungicides showed much stronger antifungal activities in YPD medium than three NFY media. Eleven fungicides inhibited growth of S. cerevisiae in all media and the other 26 fungicides showed no antifungal activity in all media. Thus, our rapid and efficient in vitro method can be considered as an alternative assay system for respiration inhibitor.

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Purification and Characterization of Urushiol Induced Laccase Isoenzyme from Fomitella fraxinea (Urushiol에 의해 유도된 장수버섯 laccase isoenzyme의 정제 및 특성)

  • Choi, Han-Seok;Park, Hyo-Suk;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.152-159
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    • 2010
  • The influence of urushiol, as an allergen on laccase property of Fomitella fraxinea was investigated. The enzyme production was reached to the highest level after 10 days, cultivation and the activity and mycelial biomass were increased by 2.5 and 1.5 folds, respectively, by adding urushiol in the culture medium. In liquid cultures using a Cu Mn-free medium, laccase lactivity was decreased by 3.8-9.2 folds, with similar dry cell weight. Two isoenzymes, were purified using anion exchange, hydrophobic interaction and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with $M_W$ around 67 kDa(Lac1) and 66 kDa(Lac2), and isoelectric points of 3.67 and 3.81. The optimal conditions for purified isoenzymes were found to be pH 4.5-5.0 and $30-35^{\circ}C$. Activity decreased by the addition of $Fe^{2+}$, $Mg^{2+}$, $Na^+$, and strongly inhibited by EDTA and sodium azide.

Scopolamine Production in Suspension Cultures of Tumor Calli from Datura metel L. (흰독말풀(Datura metel L.)종양 캘러스의 현탁배양으로부터 Scopolamine 생성)

  • 이수경;윤길영;김용해;양덕조
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.203-211
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    • 2000
  • In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

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A Review on 3D Structure Formation, Analysis and Performance Prediction Technique for All-solid-state Electrode and Battery (3차원 전고체 전극 구조체 형성, 분석 및 성능 예측 기술 동향)

  • Park, Joonam;Jin, Dahee;Kim, Dohwan;Bae, Kyung Taek;Lee, Kang Taek;Lee, Yong Min
    • Journal of the Korean Electrochemical Society
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    • v.22 no.4
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    • pp.139-147
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    • 2019
  • Lithium-ion battery (LiB) with high energy density and efficiency has been utilized for the electric vehicle (EV) and energy storage system (ESS) as well as portable devices. However, as explosion accidents have frequently happened till lately, all-solid-state lithium secondary battery (ALSB) began to get in a spotlight because it can secure a very high safety and energy density by substituting flammable organic liquid electrolyte to nonflammable inorganic solid electrolyte. In spite of ALSB's certain merits, it has shown much poorer performance of cells than one of LiB due to some challenges, which have been small or never dealt with in the LiB system. Hence, although plenty of studies made progress to solve them, an approach about design of all-solid-state electrode (ASSE) has been limited on account of difficulty of ALSB's experiments. That is why the virtual 3D structure of an all-solid-state electrode has to be built and used for the prediction of cell performance. In this study, we elucidate how to form the 3D ASSE structure and what to be needed for the simulation of characteristics on ALSB. Furthermore, the ultimate orientation of 3D modeling and simulation for the study of ALSB are briefly suggested.

Comparative Analysis of the Constituents of the Leaves and Roots of Rumex crispus and their Effects on the Differentiation of Human Osteoblast-like MG-63 Cells (소리쟁이 잎과 뿌리 성분 분석 및 사람 조골 유사 MG-63 세포 분화에 미치는 효과 비교)

  • Park, Heajin;Jeong, Jaehoon;Hyun, Hanbit;Kim, Jihye;Kim, Haesung;Oh, Hyun Il;Hwang, Hye Seong;Kim, Ha Hyung
    • YAKHAK HOEJI
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    • v.58 no.5
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    • pp.307-313
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    • 2014
  • Rumex crispus (curled dock), which is a perennial wild plant, has long been used as a laxative, astringent, and medicine to treat blood and skin diseases. We recently reported that the roots of R. crispus are an effective nutraceutical for bone. This study prepared ethanol extracts of the leaves and roots of R. crispus, and analyzed the major constituents using liquid chromatography and mass spectrometry. In addition, their effects on the proliferation and differentiation of human osteoblast-like MG-63 cells, such as cell viability, alkaline phosphatase (ALP) activity, collagen content, and mineralization, were compared. The chromatograms of the chemical constituents of the two extracts exhibited quite different profiles: quercetin and quercitrin were identified as major peaks in the leaf extract, whereas cinnamtannin B1 and procyanidin isomers were the major peaks for the root extract. Neither extract was cytotoxic at concentrations of < $25{\mu}g/ml$. ALP activity and collagen synthesis-which are markers of the early stage of osteogenesis-in MG-63 cells were significantly increased upon the addition of the root extract compared with the addition of the leaf extract. In contrast, the leaf extract had a more stimulatory effect on mineralization-which is marker of the late stage of osteogenesis-in MG-63 cells than did the root extract. In conclusion, extracts of both leaves and roots of R. crispus stimulated the bone-forming activity of osteoblasts; in particular, the root extract was more effective in the early stage of osteoblast differentiation, while the leaf extract was more effective in the late stage. This difference in anabolic activity may be due to differences in the constituents of the leaves and roots. The leaves and roots of R. crispus appear to complement each other as stimulators of bone formation.

Why did she lose her sight? A case of visual damage due to methanol inhalation (메탄올 흡입 후 발생한 시각장애 : 증례보고)

  • Han, Sangsoo;Shin, Hee-Jun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.11
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    • pp.421-425
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    • 2018
  • Methanol is a clear, colorless, volatile, and poisonous liquid that is commonly used as an industrial solvent. Visual impairment is a common symptom of methanol poisoning; however, visual impairment rarely occurs after exposure through inhalation. Therefore, visual loss after methanol intoxication via respiration has rarely been reported. We report a case of visual damage associated with methanol poisoning via respiratory exposure in an industrial setting. In this case in South Korea, a 28-year-old woman who worked at a cell phone factory was admitted to the emergency department with mental changes. She had blurred vision that began two days prior, but she did not come to the hospital until she experienced mental changes. She ranked 9 on the Glasgow Coma Scale and presented with severe metabolic acidosis. So, she was admitted to intensive care, and continuous renal replacement therapy was performed. Finally, she was discharged after recovery of her mental state, but had to undergo rehabilitation for six months. Also, her visual impairment was permanent. Methanol intoxication can occur through inhalation, which is difficult to detect initially. However, treatment of methanol poisoning is time-critical. Therefore, doctors should always keep in mind that methanol intoxication may occur via respiration. If in doubt, treatment should be given as soon as possible.

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.

Korean Red Ginseng aqueous extract improves markers of mucociliary clearance by stimulating chloride secretion

  • Cho, Do-Yeon;Skinner, Daniel;Zhang, Shaoyan;Lazrak, Ahmed;Lim, Dong Jin;Weeks, Christopher G.;Banks, Catherine G.;Han, Chang Kyun;Kim, Si-Kwan;Tearney, Guillermo J.;Matalon, Sadis;Rowe, Steven M.;Woodworth, Bradford A.
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.66-74
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    • 2021
  • Background: Abnormal chloride (Cl-) transport has a detrimental impact on mucociliary clearance in both cystic fibrosis (CF) and non-CF chronic rhinosinusitis. Ginseng is a medicinal plant noted to have anti-inflammatory and antimicrobial properties. The present study aims to assess the capability of red ginseng aqueous extract (RGAE) to promote transepithelial Cl- secretion in nasal epithelium. Methods: Primary murine nasal septal epithelial (MNSE) [wild-type (WT) and transgenic CFTR-/-], fisher-rat-thyroid (FRT) cells expressing human WT CFTR, and TMEM16A-expressing human embryonic kidney cultures were utilized for the present experiments. Ciliary beat frequency (CBF) and airway surface liquid (ASL) depth measurements were performed using micro-optical coherence tomography (μOCT). Mechanisms underlying transepithelial Cl- transport were determined using pharmacologic manipulation in Ussing chambers and whole-cell patch clamp analysis. Results: RGAE (at 30㎍/mL of ginsenosides) significantly increased Cl- transport [measured as change in short-circuit current (ΔISC = ㎂/㎠)] when compared with control in WT and CFTR-/- MNSE (WT vs control = 49.8±2.6 vs 0.1+/-0.2, CFTR-/- = 33.5±1.5 vs 0.2±0.3, p < 0.0001). In FRT cells, the CFTR-mediated ΔISC attributed to RGAE was small (6.8 ± 2.5 vs control, 0.03 ± 0.01, p < 0.05). In patch clamp, TMEM16A-mediated currents were markedly improved with co-administration of RGAE and uridine 5-triphosphate (8406.3 +/- 807.7 pA) over uridine 5-triphosphate (3524.1 +/- 292.4 pA) or RGAE alone (465.2 +/- 90.7 pA) (p < 0.0001). ASL and CBF were significantly greater with RGAE (6.2+/-0.3 ㎛ vs control, 3.9+/-0.09 ㎛; 10.4+/-0.3 Hz vs control, 7.3 ± 0.2 Hz; p < 0.0001) in MNSE. Conclusion: RGAE augments ASL depth and CBF by stimulating Cl- secretion through CaCC, which suggests therapeutic potential in both CF and non-CF chronic rhinosinusitis.

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.

Quantification of triterpenes in Centella asiatica cultivated in a smart farm, and their effect on keratinocyte activation (스마트팜 재배 병풀의 triterpenes 정량 및 각질형성세포 활성화 효과)

  • Jin Hong Park;Seong Min Jo;Da Hee Lee;Youngmin Park;Hwan Bong Chang;Tae Jin Kang;Kiman Lee
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.483-491
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    • 2023
  • This study aimed to compare the bioactive compounds in Centella asiatica (C. asiatica) cultivated in a smart farm and a field and their effects on human keratinocyte cells. C. asiatica was collected in Jeju-do, Korea, and cultured in a smart farm and a field. The main bioactive compounds in the two differentially cultured C. asiatica were identified, and their activation in keratinocytes were assessed. Amplification and sequencing of the internal transcribed spacer (ITS) DNA in the nucleus and psbA-H DNA in the chloroplast were performed for species analysis. A comparison of DNA of plants reported in the NCBI GenBank was performed. The ITS DNA and psbA-H DNA sequences of C. asiatica cultivated in a smart farm and a field were consistent with No. MH768338.1 and No. JQ425422.1, respectively. Analysis of the triterpenes was performed using high performance liquid chromatography (HPLC) and as a result, C. asiatica cultured in a smart farm had more triterpenes than those cultured in a field. The effects of C. asiatica grown in a smart farm on cell proliferation and scratch recovery in HaCaT cells were greater than those grown in a field. These results suggest that C. asiatica cultivated in a smart farm can be effectively utilized as a health functional food.