• Title/Summary/Keyword: viable cells

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Economic Production of $\gamma$-Interferon from Recombinant Human Cells in Serum Free Medium by a Moving Aeration Membrane Bioreactor (교반형 막 반응기를 이용한 재조합 인간 세포의 무혈청 배지에 의한 $\gamma$-Interferon의 생산)

  • Park, Young-Shik;Kim, Hyun-Kyu;Lim, Seo-Kyu;Park, Kyung-You;Lee, Hyeon-Yong
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.389-394
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    • 1994
  • 8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

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Cytotoxic Effect of Inonotus obliquus Composition in HCT-15 Human Colon Cancer Cells and AGS Gastric Cancer Cells (대장암 세포암종 HCT-15 세포 및 위암 세포암종 AGS 세포에서 차가버섯 조성물에 의한 세포생육 억제 효과)

  • 차재영;전병삼;문재철;유지현;조영수
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.4
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    • pp.633-640
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    • 2004
  • This study was performed to investigate the cytotoxic effect of the water-extract from Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea in HCT-15 human colon carcinoma, AGS human gastric carcinoma and NIH3T3 mouse normal fibroblast cells using viable cell count and MTT assay. The water-extract from Chaga mushroom compositions induced inhibitory effects on proliferation of HCT-15 and AGS cells in the MTT assay and viable cell count. However, mouse normal NIH3T3 cells were exhibited 80% survival under the same condition. Chaga mushroom compositions showed highly antiproliferative effect in human cancer cell line HCT-15 and AGS, but not in mouse normal cell line NIH3T3. These results suggest that Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea are the candidate for chemoprevention in colon and gastric cancer.

The Synergistic Action of the AL-Protease from Arthrobacter luteus on the Lysis of Yeast Cell Walls (Arthrobacter luteus가 생산하는 AL-Protease의 효모세포벽 용해 촉진작용)

  • Oh, Hong-Rock;Funatsu, Masaru
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.4
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    • pp.401-408
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    • 1985
  • The yeast cell wall lytic action of the alkaline AL-protease, which was found out of the crude Zymolyase that a kind of yeast cell wall lytic $endo-{\beta}-1$, 3-glucanase produced from Arthrobacter luteus, was investigated with the viable cells of S. sake and it's cell wall preparation. AL-protease on the lysis of the viable yeast cells showed very low activities with the alone, but the lytic activities were highly increased with the combination of AL-protease and Zymolyase. On the stepwise treatment of the viable yeast cells with AL-protease and Zymolyase, the cells were lysed highly only by the course having a treatment with Zymolyase after pretreatment with AL-protease. Thus synergistic action of AL-protease was not observed with any some commercial enzymes, known as a type of alkaline and serine protease such as AL-protease, and was also found to be affected greatly by the culture conditions and species of the yeast tested. AL-protease caused the release of some peptide and a lot of sugar from the cell wall preparation, but could not lysed the cell wall more than 66%. Whereas Zymolyase could lysed the cell walls almost completely with alone. On the basis of these results, the synergistic action of AL-protease on the lysis of S. sake cells is hypothesized that at first AL-protease bind to the yeast cell surface layer consisting of mannan and protein, and then changes their conformation to facilitate the penetration of Zymolyase from the outside to the inside framework layer constituted of alkali insoluble ${\beta}-1,\;3-glucan$.

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Adhesion of Kimchi Lactobacillus Strains to Caco-2 Cell Membrane and Sequestration of Aflatoxin B1 (김치 유산균의 Caco-2 세포막 부착성 및 Aflatoxin B1 제거 효과)

  • Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.5
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    • pp.581-585
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    • 2005
  • Five lactic acid bacteria (LAB) including 2 Lactobacillus strains isolated from Kimchi were evaluated to determine the binding ability to Caco-2 cells and $AFB_1$. LAB were divided into three different groups ; viable, heat-treated, and acid-treated cells. In the radioactive-labeling assay for bound cell counting, viable Lactobacillus Plantarum KCTC 3099 showed the higher adhesion to Caco-2 cells with the binding capacity of $39.2\%$, which was $149\%$ higher than Lactobacillus rhamnosus GG as a positive control. Leuconostoc mesenteroids KCTC 3100 showed the similar binding ability to L. rhamnosus GG. After 1 hour incubation at $37^{\circ}C$ with $AFB_1$, viable L. Planterum KTCC 3099 removed the toxin by $49.8\%$, which was similar level to L. rhamnosus GG. Both heat- and acid-treated groups showed high binding effect but acid-treated group was more effective for both Caco-2 cell binding and $AFB_1$ removal than the other. These results indicate that components of bacterial cell wall might be involved in tile binding to intestinal cells and toxins.

Rapid Detection of Viable Cronobacter sakazakii using Propidium Monoazide (PMA) in Combination with Real-Time qPCR (Propidium Monoazide(PMA)와 Real-Time qPCR을 이용한 살아있는 Cronobacter sakazakii의 신속검출)

  • Hwang, Dae-Geun;Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Kim, Dong-Hyeon;Song, Kwang-Young;Yim, Jin-Hyuk;Kim, Young-Ji;Kang, Il-Byung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.3
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    • pp.197-202
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    • 2015
  • While various foodborne pathogenic bacteria can be detected more rapidly via polymerase chain reaction than via conventional plating methods, it is impossible to distinguish between viable and dead cells in DNA-based assays. Hence, propidium monoazide (PMA) treatment has been introduced to detect living cells. The purpose of this study is to evaluate the applicability of the PMA treatment and real-time qPCR method for the detection of Cronobacter sakazakii and to compare it to that of plate counting. Based on our positive results, we suggest the use of PMA treatment and real-time qPCR for the detection of viable Cronobacter sakazakii in various food sources and an update of the Korean Food Code.

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The Role of Helicobacter pylori's Fur Protein in the Oxidative Stress Induced by Photodynamic Therapy (Photodynamic Therapy에 의한 산화적 스트레스 조건에서 Helicobacter pylori의 Fur 단백질의 역할)

  • Park, Yu-Na;Kim, Ji-Hoon;Choi, Sung-Sook
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.124-129
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    • 2011
  • The role of the ferric uptake regulator (Fur) of Helicobacter pylori in the oxidative stress was investigated in this study. A fur knockout mutant of H. pylori was constructed by replacing the fur gene with an aphA (kanamycin resistant marker) gene. Photodynamic therapy using methylene blue (MB) and 660 nm light was chosen to induce oxidative stress. The bactericidal effect of photodynamic therapy (PDT) was compared between wild type H. pylori and fur knockout mutant H. pylori. The degree of oxidative damage of DNA was confirmed using alkaline gel electrophoresis and an assay of 8-hydroxy-2-deoxyguanosine (8-OHdG). In control groups, the number of viable cells was maintained constantly during experiment. After PDT, the mutant H. pylori showed 10,000 times decreased viable cell number compared with wild type H. pylori. Depending on the exposure time of 660 nm light, the 3-fold increase in the concentration of 8-OHdG was observed in mutant H. pylori. The results of this study showed that H. pylori's Fur protein may play a role in oxidative stress induced by PDT.

Protective Effect of Bcl-2 in NS0 Myeloma Cell Culture is Greater in More Stressful Environments

  • Tey, B.T.;Al-Rubeai, M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.564-570
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    • 2005
  • In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.

Effect of Spirulina on Growth of Lactic Acid Bacteria (스피루리나가 유산균의 증식에 미치는 영향)

  • Son, Chan-Wok;Shin, Yu-Mi;Sim, Hyun-Jung;Kim, Mi-Yeon;Kim, Mee-Ree
    • Korean journal of food and cookery science
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    • v.23 no.6
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    • pp.968-976
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    • 2007
  • This experiment was carried out to investigate the effects of spirulina powder on the growth properties of lactic acid bacteria in reconstituted skim milk. The spirulina powder supplemented to S. thermophilus and L acidophilus slightly stimulated lactic acid production. In addition, the growth and acid production of L. bulgaricus were enhanced by the addition of spirulina powder. When the spirulina powder was added to reconstituted skim milk at the level of 1%, the mixed cultures of S. thermophilus and L. bulgaricus showed higher numbers of viable cells and higher acid production than the other cultures. The effects of the addition amounts of spirulina powder (1%, 2% and 3%) to the reconstituted skim milk on the growth properties of the mixed cultures of S. thermophilus and L. bulgaricus were evaluated. The pH of the skim milk with added spirulina powder was lower than that of the control, but the amount of spirulina did not have a significant affect. The titratable acidity increased with the incubation time until 12 hr. The number of viable cells in the skim milk with added spirulina increased according to the amount of spirulina. Thus, the spirulina was effective for the increasing lactic acid bacteria in yoghurt.

Comparative Study on Growth of Spoilage Microorganisms in Mungbean and Soybean Sprout (숙주나물과 콩나물에 대한 부패성 미생물의 증식 양상 비교)

  • 유미지;김용석;신동화
    • Journal of Food Hygiene and Safety
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    • v.19 no.1
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    • pp.25-30
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    • 2004
  • This study was conducted to investigate effect of mungbean sprout and soybean sprout against the growth of spoilage microorganism (Bacillus subtilis, Bacillus cereus, Pseudomonas fluorescens, and Listeria monocytogenes). After blanching at different conditions, the viable cells of the spoilage microorganisms were increased with storage time, but the viable cells were almost same after treating with either mungbean sprout or soybean sprout. The viable cell population in minimal broth treated with filtrate of mungbean sprout was higher than that of soybean sprout after 24 hr. However, the growth of spoilage microorganism in filtrate of mungbean sprout and soybean sprout was depending upon strain type. During incubation for 72 hr at $25^{\circ}C$, the color change of mungbean sprout was appeared moderately, but in soybean sprout it was appeared clearly at 24 hr. These results indicate that the effect of mungbean sprout and soybean sprout against growth of spoilage microorganism was not different.

In vitro antimicrobial effect of the tissue conditioner containing silver nanoparticles

  • Nam, Ki-Young
    • The Journal of Advanced Prosthodontics
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    • v.3 no.1
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    • pp.20-24
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    • 2011
  • PURPOSE. The aim of this study was to identify in vitro antimicrobial activity of the tissue conditioner containing silver nanoparticles on microbial strains, Staphylococcus aureus, Streptococcus mutans and Candida albicans. MATERIALS AND METHODS. Experimental disc samples ($20.0{\times}3.0$ mm) of tissue conditioner (GC Soft-Liner, GC cooperation, Tokyo, Japan) containing 0.1 - 3.0% silver nanoparticles (0%: control) were fabricated. Samples were placed on separate culture plate dish and microbial suspensions (100 ${\mu}L$) of tested strains were inoculated then incubated at $37^{\circ}C$. Microbial growth was verified at 24 hrs and 72 hrs and the antimicrobial effects of samples were evaluated as a percentage of viable cells in withdrawn suspension (100 ${\mu}L$). Data were recorded as the mean of three colony forming unit (CFU) numerations and the borderline of the antimicrobial effect was determined at 0.1% viable cells. RESULTS. A 0.1% silver nanoparticles combined to tissue conditioner displayed minimal bactericidal effect against Staphylococcus aureus and Streptococcus mutans strains, a 0.5% for fungal strain. Control group did not show any microbial inhibitory effect and there were no statistical difference between 24 hrs and extended 72 hrs incubation time (P > .05). CONCLUSION. Within the limitation of this in vitro study, the results suggest that the tissue conditioner containing silver nanoparticles could be an antimicrobial dental material in denture plaque control. Further mechanical stability and toxicity studies are still required.