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High Cell Density Culture of Bifidobacterium longum by Cross-flow Filtration (Cross-flow filtration에 의한 Bifidobacterium longum의 고농도 배양)

  • Lee, Myong-Suk;Park, Yun-Hee
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.18-22
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    • 1997
  • The conditions for production of high cell density of Bifidobacterium longum were investigated and the cross-flow filtration system was used to remove the inhibitory metabolites, lactic acid and acetic acid. The maximum cell growth was observed with glucose as carbon source at the concentration of 50 g/l at $37^{\circ}C$ with the initial pH 6.5. When B. longum was cultured in a cross-flow filtration system, the maximum cell growth was observed at a dilution rate(D) of $0.31\;h^{-1}$ and the dry cell weight was 16.4 g/l($3.5{\times}10^{10}\;cell/ml$), which was about four times higher than that obtained in the batch culture with pH control.

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Effect of Preoperative White Blood Cell Count on Postoperative Course in Patients with Coronary Artery Bypass Grafting (관상동맥우회술 환자에서 술 전 백혈구 수치가 수술 후 경과에 미치는 영향)

  • Son Kuk Hui;Kim Jae Ho;Kim Joung Taek;Yoon Yong Han;Kim Kwang Ho;Baek Wan Ki
    • Journal of Chest Surgery
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    • v.38 no.10 s.255
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    • pp.669-674
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    • 2005
  • Background: There are several studies that report the relationship between leukocytosis and cardiovascular disease mortality. Most of these studies stand on the basis that atherosclerosis is mediated by inflammatory process. By the same token, preoperative white blood cell count is suggested as an independent predictable factor of postoperative mortality and morbidity in coronary artery bypass grafting. The purpose of this study is to define the influence of preoperative white blood cell count on postoperative morbidity and mortality after coronary artery bypass grafting. Material and Method: The medical records of the 133 patients who had undergone isolated coronary artery bypass grafting at Inha University Hospital from 1996 to 2003 were reviewed. Patients were evenly divided into four groups, and named as group A, B, C, and D respectively based on their preoperative white blood cell count in ascending order. The number of patients in each group were 33 with exception of 34 in group A. The range of white blood cell count were from $1.3\times10^3/{\mu}L\;to\;5.9\times10^3/{\mu}L\;in\;group\;A,\; from\;6.0\times10^3/{\mu}L\;to\;7.0\times10^3/{\mu}L\;in\;group\;B,\;from\;7.1\times10^3/{\mu}L\;to\;8.9\times10^3/{\mu}L$ in group C, and from $8.9\times10^3/{\mu}L\;to\;16.9\times10^3/{\mu}L$ in group D. Result: The number of patients with recent myocardial infarction was 0 in group A, $2(6.1\%)$ in group B, $4(12.1\%)$ in group C, and $8(24.3\%)$ in D group, showing proportional increase to the white blood cell count (p<0.01). There were six postoperative deaths; $1(2.9\%)$ in group A, $1(3.0\%)$ in group B, $2(2.6\%)$ in group C, and $2(6.1\%)$ in group D (p=0.44), showing no significant difference between the groups. Postoperative wound infection occurred in 3 patients; all 3 patients were in group D, showing that postoperative wound infection is closely related to the preoperative white blood cell count. Conclusion: The association between preoperative white blood cell count and postoperative mortality could not be defined. The incidence of postoperative wound infection was found to be proportional to the preoperative white blood cell counts.

Production of Viable Lactobacillus crispatus by Using Whey Based Medium (Lactobacillus crispatus 생균 생산을 위한 whey 배지 최적화)

  • Chang, Chung-Eun;Koo, Ja-Ryong;So, Jae-Seong;Yun, Hyun-Shik
    • KSBB Journal
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    • v.26 no.6
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    • pp.529-532
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    • 2011
  • Whey based medium was optimized for the production of viable Lactobacillus crispatus KLB 46 isolated from the vagina of Korean women. Among the various nitrogen sources such as yeast extract, beef extract, and proteose peptone no. 3 supplemented to whey, beef extract showed the highest viable cell production. The addition of Tween 80 to the whey based medium increased viable cell concentration. As beef extract supplementation is not economically attractive, corn steep liquor was added as a supplementary nitrogen sources. When corn steep liquor was supplied with beef extract with the ratio 5 : 1, the viable cell count was $3.11{\times}10^9$ CFU/mL. Also, the addition of mineral salts containing sodium acetate (5 g/L), potassium phosphate dibasic (2 g/L), magnesium sulfate (0.1 g/L) and manganese sulfate (0.05 g/L) to the whey medium increased viable cell count further ($5.00{\times}10^9$ CFU/mL).

Anticancer and Immune Activities of the Extracts from Amorpha fruticosa L. (족제비싸리 부위별 추출물의 항암 및 면역활성)

  • Kim, Jung-Hwa;Kim, Dae-Ho;You, Jin-Hyun;Kwon, Min-Chul;Lee, Hyun-Jung;Lee, Hak-Ju;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.1
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    • pp.41-47
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    • 2005
  • This study was performed to experiment about useful biological activities of the parts of the extracts from Amorpha fruticosa LINNE. Experimental studies were progressed through the anticancer activities and immune activities such as cell cytotoxicity, inhibition activities of cell growth, cell growth of human B and T cell, productivity of cytokines and natural killer cell activities. The cell cytotoxicity using human Lung normal cell (HEL299) was showed cytotoxicity of below 22% by extracts of Amorpha fruticosa L. in 1.0 g/l concentration. The anticancer activities were increased in over 70% by roots extracts of Amorpha fruticosa L. in A549 and AGS cells. The immune cell growth using human immune B and T cells was increased against control by barks extracts of Amorpha fruticosa L. The secretion of cytokines $(IL-6,\;TNF-{\alpha})$ from human immune B and T cells was showed secretion of the amount of cytokines by roots extracts of Amorpha fruticosa L. Also NK cell growth was increased against control all of the extracts of Amorpha fruticosa L. From the results, the roots and barks extracts of Amorpha fruticosa L. were showed useful biological activities.

Conversion of D-$\alpha$-Amino-$\varepsilon$-Caprolactam into L-Lysine Using Cell-free Extracts of Alcaligenes eutrophus A52 (Alcaligenes eutrophus A52의 무세포 추출액에 의한 D-$\alpha$-Amino-$\varepsilon$-Caprolactam으로부터 L-Lysine으로의 전환)

  • 박희동;최선택;이인구
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.375-380
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    • 1987
  • D-$\alpha$-Amino-$\varepsilon$-carpolactam racemase (EC 5.1.1) and L-$\alpha$-amino-$\varepsilon$-caprolactam hydrolase (EC 3.5.2) were fractionated from cell-free extracts of Alcaligenes eutrophus A52 using ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. It was made sure that D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-$\alpha$-amino-$\varepsilon$-caprolactam by racemase, and then hydrolyzed into L-lysine by hydrolase in Alcaligenes eutrophus A52. For the conversion of D-$\alpha$-amino-$\varepsilon$-caprolactam into L-lysine by cell-free extracts of Alcaligenes eutrophus A52, the optimum temperature and pH were 6$0^{\circ}C$ and 8.5 respectively. The results showed that 0.5% D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-lysine at 55$^{\circ}C$ for 10 hr with a conversion rate of 98% by cell-free extracts containing 3.1mg of protein.

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Effect of Donganme (Sorghum bicolor L. Moench) against oxidative stress in vitro and in a cellular system in glial cells

  • Choi, Ji Myung;Kim, Yeo Jin;Lee, Ah Young;Cho, Eun Ju
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.497-508
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    • 2020
  • In this study, we investigated the protective effects of 'Donganme' (Sorghum bicolor L. Moench) against oxidative stress under in vitro conditions and in a cellular system using C6 glial cells. The radical scavenging activities were observed using the substrates 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl (•OH) radicals. The Donganme extract had an •OH radical scavenging activity of 82.66% at a concentration of 100 ㎍·mL-1. Additionally, when DPPH was used as the substrate, the Donganme extract exhibited a strong radical scavenging activity in a concentration-dependent manner with an IC50 value of 28.56 ㎍·mL-1. Furthermore, treating C6 glial cells with hydrogen peroxide (H2O2) reduced the cell viability and generated reactive of oxygen species (ROS) and lactate dehydrogenase (LDH) compared to the normal levels, indicating that H2O2 induced oxidative stress. However, Donganme extracts increased the cell viability and inhibited ROS and LDH production against oxidative stress by H2O2 in the C6 glial cells. In particular, it showed effective cell protection with the cell viability, ROS production, and LDH release at 83.50, 88.06, and 14.87%, respectively, which were lower than the control or similar to the normal levels even at a low concentration of 100 ㎍·mL-1. The present study suggests that the Donganme extract was effective in protecting against oxidative stress in C6 glial cells through its antioxidant activity. Thus, Donganme could be a promising therapeutic agent for neurodegenerative diseases due to oxidative stress.

The effect of KaegiBokryengHwan on sereval cancer cell lines and immuno-function (계기복령환이 수종(數種)의 암세포주(癌細胞柱) 및 면역기능(免疫機能)에 미치는 영향(影響))

  • Gang Seong-Do;Jin Cheon-Sik;Jeong Hyeon-U
    • Herbal Formula Science
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    • v.7 no.1
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    • pp.107-120
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    • 1999
  • The purpose of this Study was to investigate effects of KaegiBokryengHwan(KBH) on anti-tumor, immunocytes and nitric oxide(NO). This Study estimated the proliferation of L1210 cell lines, HeLa cell lines, SK-OV3 cell lines, MCF-7 cell lines, balb/c mouse 3T3 cell lines, mouse thymocytes and mouse splenocytes and NO production from peritoneal macrophages in vitro. and estimated the proliferation of L1210 cells, mouse thymocytes and splenocytes and NO production from peritoneal macrophages and body weight in L1210 cells-transplanted mice in vivo. The result were obtained as follow ; 1. KBH inhibited significantly SK-OV3 cell lines in vitro. 2. KBH was accelerate significantly the proliferation of balb/c mouse thymocytes in vitro. 3. KBH increased significantly NO production from peritoneal macrophages in vitro. 4. KBH didn't effect the cytotoxicity of L1210 cells in L1210 cells-transplanted mice. 5. KBH was accelerate the proliferation of splenocytes in L1210 cells-transplanted mice. 6. KBH increased NO production from peritoneal macrophages in L1210 cells-transplanted mice. 7. KBH increased the body weight as comparing with control group in L1210 cells-transplanted mice.

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In Vitro Antitumor Properties of an Isolate from Leaves of Cassia alata L

  • Olarte, Elizabeth Iglesias;Herrera, Annabelle Aliga;Villasenor, Irene Manese;Jacinto, Sonia Donaldo
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3191-3196
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    • 2013
  • Leaf extracts of Cassia alata L (akapulko), traditionally used for treatment of a variety of diseases, were evaluated for their potential antitumor properties in vitro. MTT assays were used to examine the cytotoxic effects of crude extracts on five human cancer cell lines, namely MCF-7, derived from a breast carcinoma, SK-BR-3, another breast carcinoma, T24 a bladder carcinoma, Col 2, a colorectal carcinoma, and A549, a nonsmall cell lung adenocarcinoma. Hexane extracts showed remarkable cytotoxicity against MCF-7, T24, and Col 2 in a dose-dependent manner. This observation was confirmed by morphological investigation using light microscopy. Further bioassay-directed fractionation of the cytotoxic extract led to the isolation of a TLC-pure isolate labeled as f6l. Isolate f6l was further evaluated using MTT assay and morphological and biochemical investigations, which likewise showed selectivity to MCF-7, T24, and Col 2 cells with $IC_{50}$ values of 16, 17, and 17 ${\mu}g/ml$, respectively. Isolate f6l, however, showed no cytotoxicity towards the non-cancer Chinese hamster ovarian cell line (CHO-AA8). Cytochemical investigation using DAPI staining and biochemical investigation using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-a method used to detect DNA fragmentation-together with caspase assay, demonstrated apoptotic cell death. Spectral characterization of isolate f6l revealed that it contained polyunsaturated fatty acid esters. Considering the cytotoxicity profile and its mode of action, f6l might represent a new promising compound with potential for development as an anticancer drug with low or no toxicity to non-cancer cells used in this study.

Effects of Stireed Tank Bioreactor Scale-up on Cell Growth and Alkaloids Production in Cell Cultures of Eschscholtzia californica (탱크 교반형 생물반응기의 scale-up이 Eschscholtzia californica 세포생장 및 알칼로이드 생성에 미치는 영향)

  • 유병삼;변상요
    • KSBB Journal
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    • v.13 no.6
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    • pp.700-705
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    • 1998
  • Studies were made to investigate effects of the scale-up of stirred tank bioreactors on cell growth and alkaloids production for suspension cultures of Eschscholtzia californica. In the 1.5 L STR, cell lysis was observed at 110 rpm or higher agitation speed. The agitation speed of 30 L STR was 43.7 rpm to maintain the same shear stress developed in 1.5 L STR of 100 rpm. As a result of scale-up from 1.5 L to 30 L STR, the specific growth rate was decreased from 0.12 to 0.07 day-1. The alkaloids productivity was also decreased from 0.24 to 0.14 mg/L-day. Changes of mixing performance and oxygen transfer were studied to explain the decrease of cell growth and alkaloids production. Decreased oxygen transfer rate coefficient(KLa) and increased mixing time by the scale-up was observed at various aeration rates.

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Polyhydroxyalkanoic Acid Production by Alcaligenes sp. GB-77 (Alcaligenes sp. GB-77 에 의한 Polyhydroxyalkanoic Acid의 생산)

  • 김근배;손홍주;이상준
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.220-228
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    • 1995
  • For polyhydroxyalkanoic acid (PHA) production, several microorganisms were isolated from sewage sludge. One of them, GB-77 strain, was chosen from its PHB/HV copolymer production on only fructose without cosubstrate. The isolated strain GB-77 was identified as the genus Alcaligenes. Optimal temperature and pH for cell growth were 36C and 6.8. Optimal medium composition was 10 g/l of fructose and 5 g/l of polypeptone, 1 $\times$ 10$^{-2}$M Na$^{2}$HP0$^{4}$, 1.3 $\times$ 10$^{-2}$M KH$^{2}$PO$^{4}$. To investigate the optimal condition for polyhydroxyalkanoic acid production two-stage culture technique was used; first stage for cell growth and second stage for PHA production on unbalanced growth conditions. Optimal conditions for high PHA production were C/N ratio 50, temperature 36$\circ$C and pH 6.8. To overcome fructose inhibition on cell growth, intermittent feeding fed-batch culture technique was used. Total cell concentration was 17.4 g/l with 9.1 g/l of PHA. The purified PHA was identified PHB/HV copolymer by NMR analysis.

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