• Title/Summary/Keyword: 세포분리기

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Correlation between Biosurfactants and Antifungal Activity of a Biocontrol Bacterium, Bacillus amyloliquefaciens LM11 (생물적 방제균 Bacillus amyloliquefaciens LM11의 유래 생물계면활성물질과 항균활성과의 상관관계)

  • Kang, Beom Ryong;Kim, Yong Hwan;Nam, Hyo Song;Kim, Young Cheol
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.177-185
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    • 2017
  • Bacillus amyloliquefaciens LM11 was isolated from the feces of larvae of the rhino beetle and showed strong antifungal activities against various phytopathogenic fungi by producing biosurfactants. In this study, our overall goal was to determine relationship between biosurfactants produced from the LM11 strain and its role in growth inhibition of phytopathogenic fungi. Production and expression levels of B. amyloliquefaciens LM11 biosurfactants were significantly differed depending on growth phases. Transcriptional and biochemical analysis indicated that the biosurfactants of the LM11 strain were greatly enhanced in late log-phase to stationary phase. Inhibitions of phytopathogenic mycelial growth and spore germination were directly correlated (P<0.001, R=0.761) with concentrations of the LM11 cell-free culture filtrates. The minimum inhibitory surface tension of the culture filtrate of the B. amyloliquefaciens LM11 grown in stationary phase to inhibit mycelial growth of the phytopathogenic fungi was 38.5 mN/m (P<0.001, R=0.951-0.977). Our results indicated that the biosurfactants of B. amyloliquefaciens LM11 act as key antifungal metabolites in biocontrol of plant diseases, and measuring surface tension of the cell-free culture fluids can be used as an easy indicator for optimal usage of the biocontrol agents.

Analysis of Transgenic Mouse, for the Production of Immunodeficiency Animals (면역결핍동물의 생산을 위한 형질전환생쥐의 분석)

  • 나루세겐지;양정희;이승현;최화식;이성호;박창식;진동일
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.179-185
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    • 2003
  • To determine whether the diphtheria toxin-A (DT) gene disrupts development of thymocytes in transgenic animal, the DT-A gene was used for the production of transgenic mice directed by proximal Ick promoter sequences. Two transgenic founder mice that contained several copies of transgene were produced by DNA microinjection and integration of transgene in transgenic mice was confirmed by PCR and Southern blotting analysis. Transgenic $F_1$ and $F_2$ mice were produced by outbreeding of founder and $F_1$ mice to investigate expression of transgene and phenotypes in transgneic mice. Expression of the diphtheria toxin gene was confirmed in thymus, spleen and liver of transgenic mice by RT-PCR. In circulating blood of transgenic mice, lower number of circulating white blood cells and platelets were observed compared with that of normal mice. In addition, transgneic mice had reduced number of circulating peripheral T-cells analyzed by FACS with anti-CD3 antibody. The data in these transgenic mice indicate that DT gene can play a disruptive role in developing thymocytes of transgenic mice resulted in lower number of T-cells that can be applicable to a wide range of tissues in other animals.

Ethanol Production from Tapioca Hydrolysate by Batch and Continuous Cell Retention Cultures (회분 및 연속세포유지 배양에 의한 타피오카당화액으로부터의 에탄올생산)

  • 이용석;이우기
    • KSBB Journal
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    • v.10 no.5
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    • pp.598-603
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    • 1995
  • Batch and continuous cell retention cultures were carried out using tapioca hydrolysate. In batch culture, reducing sugar of about 180g/$\ell$ was almost consumed in about 36 hours, and the concentration of ethanol produced was about 84g/$\ell$ making the ethanol yield 0.48 g-ethanol/g-(reducing sugar). The final yeast concentration was 8.5${\times}$107 cells/ml(about 2.1g/$\ell$). In a total cell retention culture operated with a dilution rate of 0.18h-1, the yeast concentration, the residual reducing sugar concentration, the ethanol concentration, and the volumetric ethanol productivity were about 40g/$\ell$, about 15g/$\ell$, 81.4g/$\ell$, and 14.7g/$\ell$-h, respectively. In another cell retention culture operated with a dilution rate and a bleed ratio of 0.2h-1 and 0.14, respectively, the yeast concentration increased to 22g/$\ell$ and the ethanol concentration oscillated around 68g/$\ell$. The volumetric ethanol productivity was about 13.6g/$\ell$-h and the residual reducing sugar concentration about 12g/$\ell$ containing glucose of about 4.5g/$\ell$. According to the results of batch fermentation using the solid residue from hydrolysate filtration as the substrate, it seemed to have a certain value. Thus, development of an effective reactor system to produce ethanol from this solid residue is in need.

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Ultrastructure of Gametes in the Three-spine stickleback, Gasterosteus aculeatus aculeatus (큰가시고기 배우자의 미세구조)

  • Deung, Young-Kun;Kim, Dong-Heui;Reu, Dong-Suck
    • Applied Microscopy
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    • v.29 no.2
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    • pp.177-187
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    • 1999
  • Ultrastructure of gametes in the three-spine stickleback, Gasterosteus aculeatus aculeatus was observed, utilizing light, scanning and transmission electron microscopes. The egg of three-spine stickleback is spherical and demersal type. The eggs are highly adhesived to each other but not to substrates. There are many oil droplets in vitelline membrane. The outer surface of egg envelope is arranged by mushroom-like structures and pore canals. The egg have a micropyle, sperm entry site, in the area of the animal pole. The egg envelope consists of three layers, an outer layer with high electron density, a middle layer consisting two layers and an inner layer consisting of 16 to 20 layers. In the fertilized egg envelope, the molecular weights of these components ranged from 14 kDa to 205 kDa. The molecular weights of nam protein bands are 19.4 kDa, 36.7 KDa, 39.4 kDa, 42.9 kDa, 46.1 kDa and 53.0 kDa. The head of spermatozoa is spherical shape and the acrosome is absent. The mitochondria in midpiece are arranged from one to three layers and separated from the axoneme by the cytoplasmic canal. The tail has two lateral fins and the axoneme is of the 9+2 structure.

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Effects of carbon source and nitrogen concentration on the P-EPS and Chl-a production at the MMBR system (MMBR에서 탄소원 종류 및 질소 농도가 S. quadricauda의 P-EPS 및 Chl-a 생성에 미치는 영향)

  • Choi, Yun-Jeong;Sim, Tae-Suk;Hwang, Sun-Jin
    • Journal of Korean Society of Water and Wastewater
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    • v.35 no.6
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    • pp.405-415
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    • 2021
  • MMBR system has been suggested as a promising system to resolve harvesting problems induced from low settling efficiency of microalgae. And recently, a lot of research on reducing fouling at the MMBR system has investigated focused on EPS in many cases. EPS of microalgae mainly consists of polysaccharides and protein components, and is produced through photosynthesis and nitrogen-carbon metabolic pathways. Especially, P-EPS is one of major compounds which occur membrane fouling phenomenon, as its hydrophobic protein components cause floc formation and cake layer accumulation. And it is already known that almost every microalgae can metabolize P-EPS or Chl-a when nitrogen sources as a substrate is insufficient or exhausted situation. With the above backgrounds, uptake rates of P-EPS or Chl-a by Scenedesmus quadricauda according to the type of carbon source and nitrogen concentration were evaluated in order to verify correlation between carbon source vs P-EPS production, and indeed Scenedesmus quadricauda uses P-EPS or Chl-a when the amounts of nitrogen sourc es in the feed is not satisfied. As a result, it was shown that P-EPS and Chl-a production were increased proportional to nitrogen concentration under organic carbon condition. And especially, the amo unts of P-EPS and Chl-a in the cell were diminished with the nitrogen source becomes insufficient or exhausted. Because P-EPS accelerates fouling at the MMBR system, P-EPS degradation by Scenedesmus quadricauda in order to get nitrogen source may contribute to reducing fouling. About a affects of N-consumed Chl-a to the MMBR fouling, more survey is needed. On the contrary, considering the purpose of MMBR system of this study, i.e. harvesting useful high value microalgae efficiently feeding adequate industrial process wastewater, it seems like difficult to maintain satisfied metabolic activity and to harvest with high yield rate using nitrogen-poor MMBR feed.

Low Temperature Inducible Acid Tolerance Response in virulent Salmonella enterica serovar Typhimurium (병원성 Salmonella enterica serovar Typhimurium의 저온 유도성 산 내성 반응)

  • Song, Sang-Sun;Lee, Sun;Lee, Mi-Kyoung;Lim, Sung-Young;Cho, Min-Ho;Park, Young-Keun;Park, Kyeong-Ryang;Lee, In-Soo
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.228-233
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    • 2001
  • The acid tolerance response (ATR) of log-phase Salmouella enterica seroyar Typhimurium is induced by acid adaptation below pH4.5 and will protect cells against more severe acid. Two distinctive ATR systems in thisorganism are a log-phase and stationary-phase ATR in which acid adaptations trigger the synthesis of acid shockproteins (ASPs). We found that log-phase ATR system was strongly affected by environmental factor, low tem-perature, $25^{\circ}C$. Exposure to low temperature and mild acid has been shown to increase acid survival dra-matically, and this survival rate was showed higher than $37^{\circ}C$. Especially unadapted cells at $25^{\circ}C$ presented tenthousand folds survival increasing when compared with cells at $37^{\circ}C$. The degree of acid tolerance of rpoSwhich is blown to be required for acid tolerance more increase than $37^{\circ}C$. Even though AIR pattern of rpoSbetween unadapted and adapted was showed similar at pH 3.1, rpoS-dependent ATR system also has beendetected in low temperature because rpoSAp prevents sustained acid survival at $25^{\circ}C$. Therefore the resultssuggest low temperature ATR system requires rpoS-dependent and -independent both. To investigate the basisfor low temperature related ATR system, gene that was participated for low temperature acid tolerance (lat) wasscreened in virulent S. enterica serovar Typhimurium UKl Using the technique of P22- MudJ (Km, lacZ)-directed lacZ operon fusion, LF452 latA‥‥MudJ was isolated. The latA‥‥MudJ of S. enterica Typhimurium pre-vented low temperature acid tolerance response. Therefore latA is considered one of the important genes for acidadaptation.

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In Vitro Properties and Biodistribution of Tc-99m and Re-188 Labeled Monoclonal Antibody CEA79.4 (Re-188과 Tc-99m 표지 단일클론항체 CEA79.4의 생체외 특성과 생체내 분포)

  • Hong, Mee-Kyoung;Jeong, Jae-Min;Yeo, Jeong-Seok;Kim, Kyung-Min;Chang, Young-Soo;Lee, Yong-Jin;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul;Lee, Seung-Jin
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.6
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    • pp.516-524
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    • 1998
  • Purpose: Radiolabeled CEA79.4 antibody has a possibility to be used in radioimmunoscintigraphy or radioimmunotherapy of cancer. We investigated the in vitro properties and biodistribution of CEA79.4 antibody labeled with Re-188 or Tc-99m. Materials and Methods: CEA79.4 was reduced by 2-mercaptoethanol to produce-SH residue, and was labeled with Re-188 or Tc-99m. For direct labeling of Tc-99m, methylene-diphosphonate was used as transchelating agent. CEA79.4 in 50 mM Acetate Buffered Saline (ABS, pH 5.3) was labeled with Re-188, using stannous tartrate as reducing agent. In order to measure immunoreactivity and the affinity constant of radiolabeled antibody, cell binding assay and Scatchard analysis using human colon cancer cells SNU-C4, were performed. Biodistribution study of labeled CEA79.4 was carried out at 1, 14 and 24 hr in ICR mice. Results: Labeling efficiencies of Tc-99m and Re-188 labeled antibodies were $92.4{\pm}5.9%$ and $84.7{\pm}4.6%$, respectively, In vitro stability of Tc-99m-CEA79.4 in human serum was higher than Re-188-CEA79.4. Immunoreactivity and affinity constant of Tc-99m-CEA79.4 were 59.2% and $6.59{\times}10^9\;M^{-1}$, respectively, while those of Re-188-CEA79.4 were 41.6% and $4.2{\times}10^9\;M^{-1}$, respectively. After 24 hr of administrations of Re-188 and Tc-99m labeled antibody, the remaining antibodies in blood were 6.32 and 9.35% ID/g respectively. The biodistribution of each labeled antibody in other organs was similar because they did not accumulate in non-targeted organs. Conclusion: In vitro properties and biodistribution of Re-188-CEA79.4 were similar to those of Tc-99m-CEA79.4. It appears that Re-188-CEA79.4 can be used as a suitable agent for radioimmunotheraphy.

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The effect of Antiseptics on the Galactolipid Metabolism of Chlorella ellipsoidea Chloroplast and Thylakoid Envelope (Chlorella ellipsoidea 엽록체막과 틸라코이드막의당지질 대사에 미치는 식품보존제의 효과)

  • 최은아;장재선;이종삼
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.221-231
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    • 1998
  • The biosynthesis of galactolipid and the composition of fatty acid in chloroplast and thylakoid envelope isolated from C. ellipsoidea treated with antiseptics (potassium sorbate: PS, sodium benzoate:SB, calcium propionate:CP) were analyzed. The contents of monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG) and total lipid in treatment with antiseptics were lower to compared with the control. The major fatty acid utilized for biosynthesis of MGDG in chloroplast envelope were palmitoleic acid (ave. 15.55%), oleic acid (ave. 15.09%) in control. Otherwise, the major fatty acids in P.S treatment were utilized for oleic acid (ave. 13.71%), linolenic acid (ave. 14.36%), palmitoleic acid (ave. 18.26%), oleic acid (ave. 17.26%) in S.B treatment, and oleic acid (ave. 16.88%), palmitoleic acid (ave. 16.31%) in CP treatment. It was showed that the major fatty acids in chloroplast envelope DGDG were oleic acid (ave. 15.75%), linolenic acid (ave. 17.74%) in control, oleic acid (ave. 14.90%), palmitoleic acid (ave. 15.97%) in P.S treatment, palmitoleic acid (ave. 13.29%), oleic acid (ave. 15.74%) in S.B treatment, and oleic acid (ave. 14.52%), palmitoleic acid (ave. 14.03%) in C.P treatment. The major fatty acid utilized for biosynthesis of MGDG in thylakoid envelope were linolenic acid (ave. 14.78%), oleic acid (ave. 12.90%) in control. Otherwise, the major fatty acids were utilized for palmitoleic acid (ave. 13.00%), palmitic acid (ave. 13.00%) in P.S treatment, palmitoleic acid (ave. 12.94%), oleic acid (ave. 12.43%) in S.B treatment, and oleic acid (ave. 12.43%), palmitoleic acid (ave. 12.43%) in C.P treatment. It was showed that the major fatty acids in thylakoid envelope DGDG were linolenic acid (ave. 18.01 %), oleic acid (ave. 15.53%) in control, linolenic acid (ave. 19.20%), linoleic acid (ave. 14.14%) in P.S treatment, palmitoleic acid (ave. 9.03%), oleic acid (ave. 14.85%) in S.B treatment, oleic acid (ave. 13.90%), linolneic acid(ave. 12.66%) in C.P treatment.

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Production of Conjugated Linoleic Acid by Lactobacillus acidophilus Isolated from Breast-Fed Infants (모유 섭취 신생아 유래 Lactobacillus acidophilus에 의한 Conjugated Linoleic Acid 생성)

  • Park, Jeong-Gyu;Song, Won-Ho;Hong, Sung-Moon;Kim, Cherl-Hyun
    • Food Science of Animal Resources
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    • v.28 no.5
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    • pp.580-586
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    • 2008
  • Conjugated linoleic acid (CLA) is a mixture of positional and geometric isomers of linoleic acid with conjugated double bonds. These conjugated dienes were found to be responsible for many biological properties related to health. The objective of this study was to evaluate the production of cis-9, trans-11 CLA by Lactobacillus acidophilus isolated from breast-fed infants. Nine different cultures were tested for their ability to produce cis-9, trans-11 CLA from free linoleic acid in MRS broth and 8% reconstituted skim milk medium supplemented with linoleic acid at $37^{\circ}C$ for 48 hr. cis-9, trans-11 CLA was not detected or detected in very small amount when cell pellets of strains grown in MRS broth and 8% reconstituted skim milk supplemented with linoleic acid of $200{\mu}g/mL$. However, free cis-9, trans-11 CLA was produced in both media. It appeared that 8% reconstituted skim milk produced more cis-9, trans-11 CLA than MRS broth. L. acidophilus NB 203 and NB 209 produced more cis-9, trans-11 CLA than other tested cultures. The inhibitory effects of supplemented linoleic acid on the growth of L. acidophilus NB 203 and NB 209 were not detected up to $3,000{\mu}g/mL$ linoleic acid addition during the growth at $37^{\circ}C$ for 48 h. The production of cis-9, trans-11 CLA by these two L. acidophilus strains increased in the logarithmic growth phase until 24 hr incubation. Under this experimental condition, the best yield of CLA isomers for L. acidophilus NB 203 and NB 209 could be obtained from medium supplemented with $500{\mu}g/mL$ linoleic acid at $37^{\circ}C$ after 24 hr of incubation. These results indicate that the use of lactic acid bacteria producing free CLA in fermented dairy products may have potential health or nutritional benefits.

Relationship Between Airway Inflammation Assessed by Induced Sputum and Airway Hyperresponsiveness in Asthmatic Patient (기관지천식 환자에서 유도객담으로 평가한 기도염증과 기관지 과민성과의 관계)

  • Chung, Lee-Young;Kim, Hyun-Jin;Jeong, Gyeong-Won;Kang, Mi-Jeong;Kim, Hwi-Jong;Chung, Hyo-Young;Lee, Jong-Deog;Hwang, Young-Sil
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.517-524
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    • 1999
  • Background : Airway inflammation and hyperresponsiveness are recognized as major characteristics of bronchial asthma. Airway inflammation has usually been assessed by invasive methods, e.g. BAL or bronchial biopsy, but recent studies proposed induced sputum as another reliable and non-invasive tool to investigate airway inflammation in asthmatic patients. Thus, the relationship between airway inflammation assessed by induced sputum and airway hyperresponsiveness was investigated in asthmatic patient. Method : Airway responsiveness was determined by the concentration that caused a 20% decrease in $FEV_1$($PC_{20}$) after inhaling incremental concentrations of methacholine. The numbers of inflammatory cells and the concentration of eosinophilic cationic protein(ECP) were assessed in induced sputum obtained by inhalation of hypertonic saline(3%). Result: We analyzed sputum induced in 15 stable asthmatic patients. The differential cell count(%) of macrophages, neutrophils, eosinophils and lymphocytes in induced sputum were $39.1{\pm}27.0%$, $29.6{\pm}21.0%$, $28.8{\pm}18.8%$, $1.3{\pm}3.1%$ respectively. The mean value of baseline FEV1(predicted) and ECP were $76.3{\pm}30.3%$ and $1,101{\pm}833{\mu}g/L$ respectively. The geometric mean value of $PC_{20}$ was 0.56 mg/mL. The relationships between the sputum eosinophil and ECP in induced sputum, and between sputum eosinophil and degree of airway responsiveness($PC_{20}$) were found to be significantly correlated (r=0.81, p<0.05 and r=-0.78, p<0.05, respectively). Sputum neutrophils and $PC_{20}$ were not correlated to each other (r=0.11, p=0.69) and a significant negative correlation was found between ECP and baseline $FEV_1$(predicted)(r=-0.62, p<0.05). Conclusion : The results of this study suggest that an induced sputum via a inhalation of hypertonic saline is useful to determine a patient's status of airway inflammation, and airway inflammation is one of the major causal factors in the development of bronchial hyperresponsiveness in asthmatic patients.

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