• Title/Summary/Keyword: 3D co-culture

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Fungicidal Efficacy of a Fumigation Disinfectant with Ortho-phenylphenol as an Active Ingredient against Trichophyton mentagrophytes, Candida albicans and Aspergillus niger (Ortho-phenylphenol을 주성분을 하는 훈증소독제의 Trichophyton mentagrophytes, Candida albicans 그리고 Aspergillus niger에 대한 살진균 효과)

  • Park, Eun-Kee;Lee, Soo-Ung;Cho, Ki-Yung;Kim, Yongpal;Yoo, Chang-Yeol;Kim, Suk;Lee, Hu-Jang
    • Journal of Environmental Health Sciences
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    • v.40 no.3
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    • pp.255-263
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    • 2014
  • Objectives: This study evaluated the fungicidal efficacy of a fumigant containing 20% ortho-phenylphenol against Trichophytone mentagrophytes (T. mentagrophytes), Candida albicans (C. albicans) and Aspergillus niger (A. niger). Methods: Five replicates of each carrier were contaminated by depositing 0.05 mL of each fungal suspension. After drying, two carriers without exposure to the fumigant and three carriers with exposure to the fumigant were left in a sealed room ($25m^3$) at $21{\pm}0.5^{\circ}C$ and $60{\pm}10%$ relative humidity for 15 hours. Immediately after removal from the test room, each carrier was transferred into recovery diluent and suspended, diluted and inoculated. After incubation, the numbers of each colony were counted, and the parameter values (N, T, d) were calculated. Results: The working culture suspension number (N value) of T. mentagrophytes, C. albicans and A. niger were $1.0{\times}10^8$, $1.2{\times}10^8$ and $5.7{\times}10^7CFU/mL$, respectively. All the colony numbers on the carriers exposed to the fumigant (n1, n2, n3) were higher than 0.5N1 (the number of fungal test suspensions by pour plate method), 0.5N2 (the number of fungal test suspensions by filter membrane method) and 0.5N1, respectively. In addition, all mean numbers of test strains recovered on the control-carriers (T value) were over $10^6CFU/mL$. For the fungicidal effect of the fumigant, all numbers of fungal reductions after exposure of the fumigant (d value) were 4 logCFU/mL. Conclusions: The present study showed that fumigant containing 20% ortho-phenylphenol has effective fungicidal activity against T. mentagrophytes, C. albicans and A. niger.

Information Flow During Individual Model Construction and Group Model Construction Type in the Sound Propagation Model Co-Construction Class (소리의 전달 모형구성 수업에서 나타난 개인모형 구성 단계 중 정보의 흐름과 모둠모형 구성의 유형)

  • Park, Jeongwoo;Yoo, Junehee
    • Journal of The Korean Association For Science Education
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    • v.38 no.3
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    • pp.393-405
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    • 2018
  • In this study, we classified the group model construction types in the actual classroom situation implementing the instructional strategy mixed with individual model construction and group model construction types. The purpose of this study is to analyze the information flow and the individual construction types of each group model construction type to obtain implications for model co-construction in a real classroom environment. A two-session class on sound propagation was carried out for thirty-three 8th grade students in Seoul. A total of 65 individual model construction and 16 group model construction processes were collected and analyzed. The group model construction types were classified as unchanged, enumerated, and elaborated. The unchanged type was found in 8 groups, the enumerated type in 3 groups, and the elaborated type in 5 groups. The isolated individual and independent construction (i.I) were found mostly in the unchanged group construction type (50.0%) and enumerated group construction type (54.5%). In the unchanged type, the radial shape of flow in which one student's information is transmitted to all the members of the group appeared. In the enumerated type, the starting point of the information flow was observed from two individuals. In the elaborated type, linear information flow appeared and both the second dissemination and reflective construction (2d.R) contributed to the group model construction (58.3%). This study suggests a viewpoint that enables to understand the process of complex model construction in an actual classroom context rather than in an ideal situation. The result of this study suggests the necessity of a modeling strategy considering the characteristics of Korean small group culture. It is expected that the discussion will progress through further studies.

In-vitro Neuroprotective Effect of Aricumin(Turmeric extract) (아리큐민의 In-vitro 신경보호 효과)

  • Yoon, Nam kyu;Kim, Byung Kwon;Ryu, Hyeon yeol;Seo, Bo Seung;Shin, Chang Ho;Kim, Kwan Kyu;Lee, Han Joo
    • The Journal of the Convergence on Culture Technology
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    • v.8 no.3
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    • pp.291-296
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    • 2022
  • This study was conducted on curcumin which had increased bioavailability as a potential AChE inhibitor for the treatment of neurodegenerative diseases. The purpose of this study is to confirm the in vitro neuroprotective effect on Aricumin (turmeric extract). To confirm the neuroprotective effect, AChE inhibition for Aricumin was evaluated, and cell viability was analyzed for HT-22cell, and oxidative stress (glutamate, H2O2)-induced HT-22 cytotoxicity was evaluated. As a result of the change in the AChE inhibition rate of Aricumin (Turmeric extract), it was confirmed that Aricumin at a concentration of 39.06㎍/ml or higher inhibited AChE activity by about 20% and more. And it was confirmed that the cytotoxicity of HT-22 cells induced by oxidative stress (Gluamate 5 mM and H2O2 500 µM) was significantly inhibited from 0.01 to 0.1 mg/ml concentration (p<005). These results suggest that Aricumin (turmeric extract) have potential neuroprotective effects.

Bactericidal Efficacy of Fumagari OPP®, Fumigant Against Staphylococcus aureus (훈증소독제, Fumagari OPP®의 Staphylococcus aureus에 대한 살균효과)

  • Cha, Chun-Nam;Park, Eun-Kee;Choi, Hyunju;Kim, Yongpal;Yoo, Chang-Yeol;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.28 no.4
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    • pp.349-353
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    • 2013
  • This study was performed to evaluate the bactericidal efficacy of Fumagari OPP$^{(R)}$, fumigation disinfectant, containing 20% ortho-phenylphenol against Staphylococcus aureus (S. aureus). In this research, efficacy test of fumigant against S. aureus was carried out according to French standard NF T 72-281. S. aureus working culture suspension number (N value), all of the colony numbers on the carriers exposed with the fumigant (n1, n2, and n3), the number of bacterial test suspentions by pour plate method (N1), the number of bacterial test suspentions by filter membrane method (N2) and the mean number of bacteria recovered on the control-carriers (T value) were obtained from the preliminary test. In addition, the reduction number of S. aureus exposed with the fumigant (d value) was calculated using T value, the mean number of bacteria in recovery solution (n'1) and the mean number of bacteria on carriers plated in agar (n'2). N value was $4.0{\times}10^8$ CFU/mL, and n1, n2, and n3 were higher than 0.5N1, 0.5N2 and 0.5N1, respectively. Additionally, T value was $3.4{\times}10^6$ CFU/mL. In the bactericidal effect of the fumigant, the d value was 6.43 logCFU/mL. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. The results indicated that Fumagari OPP$^{(R)}$ had an effective bactericidal activity against S. aureus, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with pathogenic bacteria.

Synthesis of Galactooligosaccharides in the Cheese Whey-based Medium by a Lactase from Lactobacillus paracasei YSM0308

  • Song, Tae-Suk;Lee, Kyung-Sang;Kang, Seung-Bum;Yoo, Seong-Ho;Lee, Jong-Ik;Yoon, Sung-Sik
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.565-571
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    • 2013
  • An enzyme ${\beta}$-galactosidase or ${\beta}$-galactohydrolase [EC3.2.1.23], commonly called lactase, mediates galacto-oligosaccharide (GOS) synthesis under conditions of high substrate concentrations. Also, lactase hydrolyzes ${\beta}$($1{\rightarrow}4$) lactose into glucose and galactose, the latter is successively transferred to free lactose to make various oligosaccharides via transgalactosylation. GOS is non-digestible to human digestive enzymes and has been used as a functional prebiotics. Among the 24 lactic acid bacteria (LAB) strains used, Lactobacillus paracasei YSM0308 was selected based on its exhibition of the highest ${\beta}$-galactoside hydrolysis activity, and the crude lactase was prepared for examination of reaction conditions to affect the GOS synthesis. Lactase activity was measured with a spectrophotometer using ONPG (o-nitropheyl ${\beta}$-D-galactopyranoside) method. Lactase activity was not detected in the culture supernatant and was mostly present in the cell pellet after centrifugation. Activity of the crude lactase preparation ranges from102 to 1,053 units/mL, with the highest activity determined for L. paracasei YSM0308. Optimal conditions for GOS synthesis are as follows: concentration of whey powder, pH, temperature, and time were 30%, pH 6.5-7.0, $30^{\circ}C$, and 4 h, respectively. The final GOS concentration was 19.41% (w/v) by the crude YSM0308 lactase, which was obtained from strain YSM0308 grown in the 10% (w/v) reconstituted whey-based medium.

Production and Purification of Single Chain Human Insulin Precursors with Various Fusion Peptides

  • Cho, Chung-Woo;Park, Sun-Ho;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.2
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    • pp.144-149
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    • 2001
  • For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.

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Role of Interleukin(IL)-6 in NK Activity to Hypoxic-Induced Highly Invasive Hepatocellular Carcinoma(HCC) Cells

  • Hwan Hee Lee;Hyojung Kang;Hyosun Cho
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.864-874
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    • 2023
  • Natural killer (NK) cell dysfunctions against hepatocellular carcinoma (HCC) in a hypoxic environment. Many solid tumors are present in a hypoxic condition, which changes the effector function of various immune cells. The transcription of hypoxic-inducible factors (HIFs) in cancer cells make it possible to adapt to their hypoxic environment and to escape the immune surveillance of NK cells. Recently, the correlation between the transcription of HIF-1α and pro-inflammatory cytokines has been reported. Interleukin (IL)-6 is higher in cancers with a highly invasive ability, and is closely related to the metastasis of cancers. This study showed that the expression of HIF-1α in HCC cells was associated with the presence of IL-6 in the environment of HCC-NK cells. Blocking of IL-6 by antibody in the HCC-NK interaction changed the production of several cytokines including TGF-β, IL-1, IL-18 and IL-21. Interestingly, in a co-culture of HIF-1α-expressed HCC cells and NK cells, blocking of IL-6 increased the production of IL-21 in their supernatants. In addition, the absence of IL-6 significantly enhanced the cytotoxic ability and the expression of the activating receptors (NKG2D, NKp44, and NKG2C) in NK cells to HIF-1α-expressed HCC cells. These effects might be made by the decreased expression of HIF-1α in HCC cells through the inhibited phosphorylation of STAT3. In conclusion, the absence of IL-6 in the interaction of HIF-1α-expressed HCC cells and NK cells could enhance the antitumor activity of NK cells to HCC cells.

Secondary Metabolites from the Mycelial Culture Broth of Phellinus linteus (상황(Phellinus linteus) 배양 균사체의 2차 대사산물에 대한 화학적 연구)

  • Song, K.S.;Cho, S.M.;Ko, K.S.;Han, M.W.;Yoo, I.D.
    • Applied Biological Chemistry
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    • v.37 no.2
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    • pp.100-104
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    • 1994
  • From the 48 hour-cultured mycelial broth of Phellinus linteus, six compounds were isolated by means of ethyl acetate extraction, silica gel column chromatography and preparative thin layer chromatography, consecutively. Compound 1 was identified as a succinic acid by the comparison of its spectral data with authentic sample. Compounds 2 and 3 were identified as p-hydroxyphenyl acetic acid methyl ester and p-hydroxybenzaldehyde by spectroscopic studies, respectively. NMR and MS studies of compound 6 revealed that it was 2,5-dihydroxymethyl furan. Compound 4, which showed similar NMR absorptions and MS fragmentation pattern with those of compound 6 was identified as 2-hydroxymethyl-5-methoxymethylfuran. These structures were verified by the spectral data of the acetate derivatives of the compounds. Compound 5 was supposed to be a N-acetyltyramine from its $^1H-NMR$ and EI-MS data, and its structure was confirmed by a synthesis starting from tyramine.

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Production of Poly(Hydroxybutyric-Co-Hydroxyvaleric) Acid by Pseudomonas sp. HJ (Pseudomonas sp. HJ에 의한 Poly(Hydroxybutyric-Co-Hydroxyvaleric) Acid의 생산)

  • 손홍주;민관필이상준
    • KSBB Journal
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    • v.10 no.4
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    • pp.349-356
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    • 1995
  • To produce PHA(polyhydroxyalkanoic acid) from microbr, dozens of microorganism have been screened from sewage sludge. Selected a strain HJ out of 50 strains of PHA producing bacteria has a capability of accumulating large amounts of PHB/HV copolymer when grown in batch culture with a single carbon source (glucose) that was not generally considered as precursor of hydroxyvalerate monomer unit. The strain HJ was identified as the genus Pseudomonas with respect to morphological, cultural, and biochemical characteristics. The optimal temperature and pH for cell growth were $37^{\circ}C$ and 7.0. The optimal medium compositions for cell growth were glucose 1% as a carbon source, (NH4) 2SO4 0.2% as a nitrogen source, K2HPO4 0.3%, and KH2PO4 0.45%. TO investigate she optimal condition for PHA production two-step cultivation method was employed. PHA production was inducted by deficiency of NH4+, SO4-2, Mg+2. Besides carbon source, deficiency of all nutrients stimulated PHA productivity but deficiency of NH4+ stimulated the most HV monomer content. The highest PHA production was C/N molar ratio 95.2. Pseudomonas sp. HJ was also able to pyoduc PHB/HV copolymer when cultivated on alkane, alkanoate, alcohol as carbon sources. The contents of PHA and she proportions of hydroxyvalerate monomer units varied depending on the carbon sources. Especially Pseudomonas sp. HJ was able to incorporate hydroxyvalerate into PHB/HV to level as high as from 49 to 74 mol% when grown in a medium containing hexadecane and propionate. The purified PHA was identified PHB/HV copolymer by HNMR analysis.

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Cigarette Smoke Extract-Treated Mouse Airway Epithelial Cells-Derived Exosomal LncRNA MEG3 Promotes M1 Macrophage Polarization and Pyroptosis in Chronic Obstructive Pulmonary Disease by Upregulating TREM-1 via m6A Methylation

  • Lijing Wang;Qiao Yu;Jian Xiao;Qiong Chen;Min Fang;Hongjun Zhao
    • IMMUNE NETWORK
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    • v.24 no.2
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    • pp.3.1-3.23
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    • 2024
  • Cigarette smoke extract (CSE)-treated mouse airway epithelial cells (MAECs)-derived exosomes accelerate the progression of chronic obstructive pulmonary disease (COPD) by upregulating triggering receptor expressed on myeloid cells 1 (TREM-1); however, the specific mechanism remains unclear. We aimed to explore the potential mechanisms of CSE-treated MAECs-derived exosomes on M1 macrophage polarization and pyroptosis in COPD. In vitro, exosomes were extracted from CSE-treated MAECs, followed by co-culture with macrophages. In vivo, mice exposed to cigarette smoke (CS) to induce COPD, followed by injection or/and intranasal instillation with oe-TREM-1 lentivirus. Lung function and pathological changes were evaluated. CD68+ cell number and the levels of iNOS, TNF-α, IL-1β (M1 macrophage marker), and pyroptosis-related proteins (NOD-like receptor family pyrin domain containing 3, apoptosis-associated speck-like protein containing a caspase-1 recruitment domain, caspase-1, cleaved-caspase-1, gasdermin D [GSDMD], and GSDMD-N) were examined. The expression of maternally expressed gene 3 (MEG3), spleen focus forming virus proviral integration oncogene (SPI1), methyltransferase 3 (METTL3), and TREM-1 was detected and the binding relationships among them were verified. MEG3 increased N6-methyladenosine methylation of TREM-1 by recruiting SPI1 to activate METTL3. Overexpression of TREM-1 or METTL3 negated the alleviative effects of MEG3 inhibition on M1 polarization and pyroptosis. In mice exposed to CS, EXO-CSE further aggravated lung injury, M1 polarization, and pyroptosis, which were reversed by MEG3 inhibition. TREM-1 overexpression negated the palliative effects of MEG3 inhibition on COPD mouse lung injury. Collectively, CSE-treated MAECs-derived exosomal long non-coding RNA MEG3 may expedite M1 macrophage polarization and pyroptosis in COPD via the SPI1/METTL3/TREM-1 axis.