• Title/Summary/Keyword: agar-free

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Presence of Leukemia-maintaining Cells in Differentiation-resistant Fraction of K562 Chronic Myelogenous Leukemia (만성 골수성 백혈병 K562세포의 분화 내성 분획에서 백혈병 유지 세포의 동정)

  • Lee, Hong-Rae;Kim, Mi-Ju;Ha, Gahee;Kim, So-Jung;Kim, Sun-Hee;Kang, Chi-Dug
    • Journal of Life Science
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    • v.23 no.2
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    • pp.197-206
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    • 2013
  • The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.

Isolation and Identification of Lactic Acid Bacteria with Probiotic Activities from Kimchi and Their Fermentation Properties in Milk (전통 김치로부터 Probiotic 유산균의 분리 및 우유 발효 특성)

  • Lim, Young-Soon;Kim, JiYoun;Kang, HyeonCheol
    • Journal of Dairy Science and Biotechnology
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    • v.37 no.2
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    • pp.115-128
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    • 2019
  • Lactic acid bacteria obtained from traditional Kimchi were selected on the basis of their caseinolytic activity and lactose usability and examined for availability as a starter in probiotic activity. Thirty-two strains were selected as lactic acid producing bacteria in BCP agar, and two strains (KC23 and KF26) with more than 90% resistance for both acid and bile salts were selected. The two strains were identified as L. plantarum (KC23) and L. paracasei (KF26) by API 50 CHL system and 16S rRNA sequence analysis. L. plantarum (KC23) was finally selected based on its biochemical characteristics for lactose and raffinose usability. Free tyrosine content increased rapidly in 10% skimmed milk medium, from $24.1{\mu}g/mL$ after 8 h to $43.9{\mu}g/mL$ after 16 h. Additionally, the caseinolytic clear zone of 12 mm of L. plantarum (KC23) was greater than the 9 mm zone of commercial L. acidophilus CSLA. The bacterium exhibited mesophilic growth and yielded $8.9{\times}10^8CFU/mL$ when incubated at $37^{\circ}C$ for 12 h at pH 4.25. Moreover, L. plantarum KC23 exhibited antibacterial activity as it formed a clear zone of 8-13 mm for the 5 pathogens. Adherent activity was 2.23 fold higher than that of LGG. The acidity of 10% skimmed milk fermented for 12 h was 0.74%.

Antioxidant and Antimicrobial Activities of Fruiting Bodies of Phellinus gilvus Collected in Korea (국내에서 수집된 마른진흙버섯 자실체의 항산화 및 항균 효과)

  • Yoon, Ki-Nam;Jang, Hyung Seok
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.4
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    • pp.355-364
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    • 2016
  • This study was initiated to evaluate the antioxidant and antimicrobial activities of methanol extract (ME) and hot water extract (HWE) obtained from the fruiting bodies of medicinal mushroom, Phellinus gilvus. The free radical scavenging activity of ME from P. gilvus on 1,1-diphenyl-2-picrylhydrazyl (DPPH) were 93.65% at 2 mg/mL, which was comparable with the positive control, butylated hydroxytoluene (BHT, 96.97%) at the same concentration. The ferrous ion-chelating ability of ME and HWE was significantly higher than that of BHT at all concentration levels. The antimicrobial assay of ME was performed against six bacteria and one species of fungus. ME exhibited antibacterial activity against 5 out of 6 bacteria: Staphylococcus aureus, Streptococcus mutans, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa; whereas, ME did not show antimicrobial activity against gram-negative bacterium Vibrio vulnificus and fungal species Candida albicans. The minimum inhibitory concentration (MIC) of ME against 5 strains of bacteria, such as S. aureus, S. mutans, B. subtilis, E. coli, and P. aeruginosa, was 100, 100, 50, 100, 200 mg/mL, respectively. The results suggest that good antioxidant and microbial activities of P. gilvus fruiting bodies might be used for natural antioxidant and antimicrobial agents.

Crystal Structure and Fungicidal Activity of N-[1-(benzotriazol-1-yl)alkyl]aniline Derivatives (N-[1-(benzotriazol-1-yl)alkyl]aniline 유도체의 결정구조와 항균활성)

  • Lim, Chi-Whan;Yi, Kyeong-Joon;Sung, Nack-Do
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.359-363
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    • 1995
  • New ten N-[1-(benzotriazol-1-yl)alkyl]aniline(4) derivatives were synthesized and the crystal structure of 4h was shown by X-ray crystallography and the absolute configuration has been assigned as S form. The molecule crystallizes in the monoclinic system, space group $P2_{1}/n$. And the molecules in the crystal are linked with each other through the hydrogen bond $(N_{11}-H_{11}{\cdots}N3)$ with distance $2.300(11){\AA}$ The fungicidal activity($pI_{50}$) in-vitro against Botrytis cineria (BC), Phytophthora casici (PC) and Sclerotium cepinorum (SC) were determined by the agar dilution method. The structure activity ralationships (SAR) between structure of 4 and the activity were studied using a physicochemical parameters of substituents and multiple regression technique. Among these compounds, only the bromo group substituent(4f) showed higher activity, which depend on the hydrophobic(${\pi}$) of substituents. The relative orders of the activity are SC>BC> and PC, respectively. This implies that the activity is affected by the hydrophobic(${\pi}$) nature of the Z group rather than the X group. Linear free energy relationships(LFER) on the fungicidal activity with substituents has been also discussed.

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Detection and Distribution of the Pathogenic Bioagent Aeromonas (Gamma-Proteobacteria) in Water Supplies of Seoul (서울시 상수계통에서 병원성균 Aeromonas (감마-프로테오박테리아) 분포연구)

  • Lee, Eun-Sook;Lee, Mok-Young;Han, Sun-Hee;Ka, Jong-Ok
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.106-110
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    • 2007
  • The detection and distribution of Aeromonas in water supplies were investigated by using the USEPA Method 1605. Water samples were collected from the Han River, finished waters and tap waters supplied from Water Treatment Plants in Seoul monthly from July 2002 to December 2003. Aeromonas species in each water sample were quantified based on the development of yellow colonies on the surface of membrane filter using a selective medium (Ampicillin-Dextrin Agar with Vancomycin). The Quality Control (QC) for this study met the acceptance criteria of Method 1605. The concentrations of Aeromonas species in surface water samples ranged from $1.0{\times}10^{0}\;to\;9.8{\times}10^{3}\;CFU/ml$. Aeromonas species were found only in one tap water sample with concentration of 1 CFU/500 ml. No Aeromonas species were found in any finished water samples. Aeromonas species detected here were identified as A. salmonicida(51%), A. caviae(4.7%), A. schubertti(3.4%), A. sobria(3.8%), A. hydrophila(2.1%), and A. ichithiosmia(0.4%). A. salmonicida was the dominant species, which is of no significance to human health. Chlorine resistance of A. salmonicida was evaluated and as a result, 99.99% of A. salmonicida decreased after 30 seconds exposure at residual free chlorine 0.2 mg/L. These suggest that the waters supplied in Seoul may be safe against the pathogenic agent Aeromonas.

Studies on the EPA(eicosapentaenioc acid) production by marine bacteria (해양세균(海洋細菌)을 이용(利用)한 EPA(eicosapentaenoic acid) 생성(生成)에 관한 연구(硏究))

  • Joh, Yong-Goe;Kim, Seong-Jin;Kim, Ji-Soo;Lee, Min-Kyeng
    • Journal of the Korean Applied Science and Technology
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    • v.12 no.2
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    • pp.121-130
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    • 1995
  • This project has been worked out for isolation of EPA-producing bacteria from marine source of sea water, sea sediment and intestinal contents eviscerated from some red-muscle fish such as mackerel, horse-mackerel and spike fish. The samples were precultured on the media of PPES-II glucose broth and then pure-cultured on Nutrient agar and P-Y-M glucose. Lipids extracted from those bacterial mass collected by centrifugation were analysed in terms of lipid class and fatty acid composition. The results are resumed as follows : 1. 112 strains from sea water and 76 strains from sea sediment were tested for their EPA producing capability, but both strains of (SA-67 and SA-91) from the former and four strains(SS-35, 37, 51 and 71) from the latter have been proved to produce EPA above the level of 2% of total fatty acids. The strains such as GS-11, 29, 31, HM-9, 29, B-18, 33, 107, YL-129, 156, 203, 77, 104 and 256 which were isolated from fish intestinal contents, have also produced EPA at higher level than 2% of total fatty acids. 2. Contents of total lipids extracted from the cultures of these strains grown at $25^{\circ}C$, range from 2.8% to 6.9% (on dry weight %), and they are mainly composed of polar lipids($40.9{\sim}52.9%$) such as phosphatidyl glycerol($^{+}cardiolipin$)(?) and phosphatidyl ethanolamine ($33.8{\sim}40.0%$), with smaller amount of free fatty acid ($11.2{\sim}20.2%$). 3. EPA was isolated from a mixture of fatty acid methyl esters obtained from the lipid of each strain by HPLC in silver-ion mode and was identified by GC-Mass spectrometry. 4. The strains of SW-91, GS-11, GS-29, HM-9, B-18 and YL-203 grown at $25^{\circ}C$ have a level of 5% EPA in their total fatty acids, and the GS-11 and HM-9 strains show a tendency of increase in the EPA level with an increase of growth temperature.

Biochemical Characteristics of Lactobacillus acidophilus Isolated from a Breast-Fed Infant (모유 섭취 신생아 유래 Lactobacillus acidophilus의 생리적 특성)

  • Hong, Sung-Moon;So, Byung-Chun;Yoon, Seumg-Won;Kim, Cheol-Hyun
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.1
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    • pp.45-53
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    • 2012
  • Lactobacillus acidophilus isolated from the feces of a 7-day-old breast-fed infant was characterized to examine the scope of its commercial use. Forty-three Lactobacillus strains, which could grow at pH 5.5, were isolated. From these Lactobacillus isolates, 14 Lactobacillus strains were selected, which demonstrated more than 80% viability and homofermentative lactic fermentation. Finally, 9 L. acidophilus strains (NB 201~NB 209) were identified as candidate strains based upon biochemical properties, carbohydrate utilization, and cellular fatty acid composition. L. acidophilus isolates demonstrated a survival rate of more than 80% when exposed to pH 2.5 for 2 h. In particular, L. acidophilus NB 204 showed a strong acid tolerance, with a 71% survival rate even at pH 2.0. L. acidophilus isolates also manifested high bile acid tolerance; more than 87% of the cells survived on agar containing 1% bile extract, except for L. acidophilus NB 206, which showed a 73% survival rate. All L. acidophilus isolates were confirmed to have proteolytic activity; L. acidophilus NB 204 and NB 209 yielded higher levels of TCA-soluble peptides and free amino acids. The ${\beta}$-galactosidase activity of the L. acidophilus isolates was in the range of 1.97~2.45 units/mL.

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Antimicrobial Effects and Antioxidative Activities of the Cosmetic Composition Having Natural Plant Pigments (천연색소 함유 화장료 조성물의 항균효과 및 항산화활성)

  • Boo, Hee-Ock;Shin, Ji-San;Hwang, Sung-Jin;Bae, Chun-Sik;Park, Su-Hyun
    • Korean Journal of Plant Resources
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    • v.25 no.1
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    • pp.80-88
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    • 2012
  • This study was conducted to clarify the antimicrobial effects and radical scavenging activities of the cosmetic compositions having the natural plant pigments, and to enhance the natural materials utilization of cosmetics. The antimicrobial activities of the fifteen kinds of cosmetic composition having natural plant pigments were evaluated using the agar diffusion method. Most of the cosmetic composition having the natural pigments showed the clear zone formation of growth inhibition against Bacillus subtilis, Staphylococcus aureus, Listeria monocytogenes, Vibrio parahaemolyticus and Aspergillus flavus. Especially, purple sweet potato, bitter melon, mulberry leaf and gromwell showed the higher antimicrobial activities. All the cosmetic compositions were evaluated for their antioxidant activity using DPPH radical scavenging capacity and nitrite scavenging ability activity. Both of the free radical DPPH and nitrite scavenging ability was the highest in the cosmetic compositions of onion peel, and these antioxidant activity was significant differences according to different plant pigments. In this study, we conjectured that the plant pigment had the potent biological activities, therefore these plant resources having functional components could be a good materials for development into source of natural cosmetics.

Isolation and Identification of Agarose-degrading Bacterium, Pseudoalteromonas sp. GNUM08122 (아가로오스 분해세균인 Pseudoalteromonas sp. GNUM08122 분리 및 동정)

  • Kim, Yu-Na;Jeong, Yeon-Kyu;Kim, Mu-Chan;Kim, Sung-Bae;Chang, Yong-Keun;Chi, Won-Jae;Hong, Soon-Kwang;Kim, Chang-Joon
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.1-9
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    • 2012
  • This study's aim was to isolate microorganisms producing agarase with a high activity, with possible applications in improving the performance of the pretreatment processes for bioethanol production. Marine algaes were collected from the south coast of Korea, from which three kinds of microorganisms were isolated. After a 4-day culture of these strains at $25^{\circ}C$, crude enzymes were obtained from culture supernatant or cell-free extract by ammonium sulfate precipitation and membrane dialysis. Agarase activity was observed in these crude enzymes. Notably higher specific activity was observed in the crude enzyme obtained from the culture supernatant rather than that from the cell-free extract. This indicates that a secreted enzyme has a much greater activity than a cellular enzyme. Crude enzymes from the GNUM08122 strain were inferred to have ${\alpha}$-agarase activity because release of p-nitrophenol was observed, possibly due to the cleavage of p-nitrophenyl-${\alpha}$-D-galactopyranoside. The 16S rRNA sequence of GNUM08122 showed a close relationship to Pseudoalteromonas issachenkonii KMM 3549 (99.8%) and Pseudoalteromonas tetraodonis IMA 14160 (99.7%), which led us to assign it to the genus Pseudoalteromonas. Biochemical and physiological study revealed that this strain can grow well at $40^{\circ}C$ under a wide range of pH (pH 4~8) in high-salt conditions (10% NaCl).

Manufacture of Cutting-Gouda Cheese using Lactobacillus rhamnosus_p1 and the Physicochemical Properties of Gouda Cheese during Ripening Periods (Lactobacillus rhamnosus_p1을 이용한 절단형 고다치즈 제조방법 및 숙성 중 품질특성)

  • Park, Jong-Hyuk;Jung, Hoo-Kil;Moon, Hye-Jung;Oh, Jeon-Hui;Lee, Joo-Hee;Kim, Myung-Kon;Na, Sang-Eon;Kim, Youn-Jeong;Hwang, Young-Tae
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.131-139
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    • 2014
  • The aim of this study was to manufacture Cutting-Gouda cheese and to investigate the change in physicochemical properties of Cutting-Gouda cheese made with Lactobacillus rhamnosus_p1. Lactic acid bacteria were isolated from Gouda cheese ripened for more than 1 year. They were identified as 2 strains of L. rhamnosus, Lactobacillus casei, Lactobacillus curvatus, and Staphylococcus saprophyticus by 16S rDNA sequencing and named L. rhamnosus_p1, L. casei_p2, L. curvatus_p3, L. rhamnosus_p4 and S. saprophyticus_p5. The proteolytic activities of isolated strains against casein were measured using prepared skim milk agar plates. L. rhamnosus_p1 showed the highest proteolytic activity. Cutting-Gouda cheese was made with L. rhamnosus_p1, and its physicochemical properties (moisture, protein, fat, ash and free amino acid content) were measured during ripening periods. Because of the modified atmosphere packaging ($N_2{^-}$), there was no change in moisture, protein, fat, and ash in the experimental group. The total amount of free amino acids in the control and experimental group gradually increased during ripening periods. The sensory evaluation showed that the experimental group was preferable to the control group. This result suggests that L. rhamnosus_p1 has potential to be developed as a new starter for Gouda cheese.

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