• Title/Summary/Keyword: cell Iysis

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Preventive Effect of Lysis in the Cadmium-Tolerant Hansenula anomala B-7 Cells by Metal Ions (금속 이온에 의한 카드늄 내성균주 Hansenula anomala B-7 세포의 용균 방지 효과)

  • Song, Hyung-Ik;Yu, Tae-Shick
    • The Korean Journal of Mycology
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    • v.19 no.4
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    • pp.282-284
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    • 1991
  • The yeast, Hansenula anomala B-7, isolated from the $Cd^{2+}$ rich soils and determined to be tolerant in the high concentration of $Cd^{2+}$ were employed in this work. Its intact cells grown in high concentration of $Cd^{2+}$ were observed to be Iysed at the early stage when transferred to a cadmium deficient broth. Its intact cells found to be not Iysed and grow well under the high concentration of $Cd^{2+}$. The Iysis of the intact cells grown at the high concentration of $Cd^{2+}$ ion was not found when the metal ions were replaced with $Cd^{2+}$ ion in the same concentration. This result indicated that Iysis of yeast cells, at least in this isolate, would be related to cell osmosis with the mineral ions added.

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Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
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    • v.3 no.1
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    • pp.73-78
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    • 1999
  • Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

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Mode of Action of the Bacteriocan from Lactobacillus sp. GM7311 against Gram Negative Bacteria (Lactebacillus sp. GM7311이 생산하는 박테리오신의 Gram 음성균에 대한 작용형태)

  • KANG Ji-Hee;LEE Myung-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.2
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    • pp.139-143
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    • 1999
  • The antimicrobial action of bacteriocin produced by lactobacillus sp. GM7311 against three Gram negative bacteria, Proteus mirabilis, Vibrio parahaemolyticus, and Escherichia coli, was investigated, When the bacteriocin was added to the culture at different stages, viable cells of all of the indicator strains tested were decreased, even though the most inhibited indicator cell growth stages were different. Transmission electron microscopic observation of indicator strains treated with bacteriocin revealed cell Iysis, indicating the cell membrane appears to be the primary site of action. The amino acids concentration of indicator strains treated with bacteriocin were diminished and fatty acids compositions were changed as compared with controls.

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Mode of Action of the Bacteriocin from Lactobacillus sp. GM7311 against Gram Positive Bacteria (Lactobacillus sp. GM7311이 생산하는 박테리오신의 Gram 양성균에 대한 작용형태)

  • KANG Ji Hee;LEE Myung Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.4
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    • pp.560-566
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    • 1998
  • The bacteriocin produced by Lactobacillus sp. GM7311 showed strong inhibitory activity against the growth of three Gram positive bacteria, Listeria monocytogenes, Bacillus subtilis, and Staphylococcus aureus. When the bacteriocin was added to the culture at different phases, viable cells of all of the tested strains were decreased, although the most inhibited phase was different. Thereby, when the bacteriocin $(100\;Bu/m{\ell})$ was added to exponential and stationary phase of L. monocytogenes, the rapid reduction of viable cell counts occured. And, in the case of B. subtilis, the highest inhibitory effect occured at lag phase and mid-exponential phase by the addition of the bacteriocin under same condition as mentioned above. Also, we can observe the accelerated reduction of survivors counts for the all of the phase except stationary phase in the S. auresus. Transmission electron microscopic observation of L. monocrogenes and B. subtilis treated with bacteriocin revealed apparent Iysis of the cell wall and excretion of the cell contents, indicating bacteriolysis. Also, the amino acids and fatty acids compositions were different from controls. However, the Iysis of cell wall didn't occur in S. aureus, though the cytoplasmic materials were reduced. This result indicates that the bacteriocin inhibits the synthesis of nuclear materials such as DNA, RNA and proteins.

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Effect of Arsenic on Heat Shock Protein and Vascular Contractility of Rat Aorta (횐쥐 대동맥의 수축반응과 열충격단백질에 대한 비소의 영향)

  • 박태규;권윤정;김중영
    • Journal of Environmental Science International
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    • v.12 no.6
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    • pp.651-657
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    • 2003
  • In order to examine if arsenic, one of environmental stresses, contributes to hypertension as one of cardiovas cular pathological factors, this study was perfarmed in vivo and in vitro, using intacted or pithed rats and aorta ring preparation, respectively. And also the relationship between expression of heat shock protein (HSP) 90 and vasoactives-induced contractile response was elucidated. To measure blood pressure, the carotid arterial pressure was recorded on physiograph(Grass Co. 79E) connected to strain gauge. On the other hand, contractile response of vascular ring preparation isolated from rat was determined in organ bath and was recorded on physiograph connected to isometric transducer. And HSP was detacted by Western blotting whole cell Iysis. Preganglionic nerve stimulation was increased by 26.0% in arterial pressure of rat treated with arsenic. Vascular contractile response was monitored and HSP were measured by Western blotting of whole Iysis prepared from samples exposed with 0, 0.5, 1, 2 and 4 mM of arsenic for 8 hours. The dose-vascular responses of potassium chloride were augmented by increasing dose of arsenic in the strips exposed to arsenic for 8 hours, and were not augmented for 1, 3, 5 hours. And the response of relaxation of rat aorta induced by histamine was not influenced by arsenic stress. The increase of HSP 90 expression in rat aorta was pronounced at 8 hours after 4 mM of arsenic treatment, but HSP 60 expression was not. Arsenic stress not only increased the expression of HSP 90 in the rat aorta, but also augmented contractions to potassium chloride. These results indicated that arsenic stress was sufficient to induce heat shock protein 90, resulting in increased vascular contractility in rat aorta.

Effect of irradiation on the Streptococcus mutans (방사선조사가 Streptococcus mutans에 미치는 영향)

  • Ahn, Ki-Dong;Kim, Gyu-Tae;Choi, Yong-Suk;Hwang, Eui-Hwan
    • Imaging Science in Dentistry
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    • v.37 no.1
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    • pp.35-43
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    • 2007
  • Purpose : To observe direct effect of irradiation on cariogenic Streptooccus mutans. Materials and Methods : S. mutans GS5 was exposed to irradiation with a single absorbed dose of 10, 20, 30, and 40Gy. Viability and changes in antibiotic sensitivity, morphology, transcription of virulence factors, and protein profile of bacterium after irradiation were examined by pour plate, disc diffusion method, transmission electron microscopy, RT-PCR, and SDS-PAGE, respectively. Results : After irradiation with 10 and 20Gy, viability of S. mutans was reduced. Further increase in irradiation dose, however, did not affect the viability of the remaining cells of S. mutans. Irradiated 5. mutans was found to have become sensitive to antibiotics. In particular, the bacterium irradiated with 40Gy increased its susceptibility to cefotaxime, penicillin, and tetracycline. Under the transmission electron microscope, number of morphologically abnormal cells was increased as the irradiation dose was increased. S. mutans irradiated with 10 Gy revealed a change in the cell wall and cell membrane. As irradiation dose was increased, a higher number of cells showed thickened cell wall and cell membrane and Iysis, and appearance of ghost cells was noticeable. In RT-PCR, no difference was detected in expression of gtfB and spap between cells with and without irradiation of 40Gy. In SDS-PAGE, proteins with higher molecular masses were gradually diminished as irradiation dose was increased. Conclusion : These results suggest that irradiation affects the cell Integrity of S. mutans, as observed by SDS-PAGE, and as manifested by the change in cell morphology, antibiotic sensitivity, and eventually viability of the bacterium.

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Cholesterol Uptake by Lactobacillus acidophilus: Its Fate and Factors Influencing the Uptake (Lactobacillus acidophilus에 의한 콜레스테롤의 흡착)

  • Noh, Dong-Ouk
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.2
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    • pp.195-206
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    • 1996
  • Cholesterol assimilated by Lactobacillus acidophilus ATCC 43121 was not metabolically degraded in that most of it was recovered with the cells. Cells grown in the presence of cholesterol micelles and bile salts were more resistant to Iysis by sonication than those grown in their absence, suggesting a possible alteration of cellular membranes. Cholesterol assimilation occurred during growth at pH 6.0, the amount of which was more than that by cells grown without pH control. Cholesterol assimilated by cells was recovered in the membrane fractions of cells both grown at pH 6.0 and without pH control. The effect of unsaturated fatty acids on cholesterol assimilation was not clear, since there was no significant (P> 0.05) difference in the amount taken up from micelles prepared using L-${\alpha}$-phosphatidylcholine, dioleoyl or L-${\alpha}$-phosphatidylcholine, distearoyl. Without Tween 80, little, if any, cell growth or cholesterol uptake was observed. In the presence of 0.05% Tween 80, cholesterol uptake increased dramatically as did growth. However, as the amount of Tween 80 increased beyond 0.05%, cholesterol uptake decreased while the amount of growth remained the same.

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Isolation and Characterization of Algicidal Bacteria KY1 (살조세균 KY1의 분리와 특성조사)

  • PARK Keun-Young;KIM Mi-Ryung;KIM Sung-Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.452-457
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    • 1999
  • Algicidal bacteria were isolated from the mud of south coastal sea of Korean peninsula and screened to evaluate algicidal activity on Cochlodinium polykrikoides. The optimum condition for the development of highest algicidal activity was determined, The optimum sodium chloride concentration for algicidal activity of isolated algicidal bacteria was $3\%$. The optimum temperature, pH and culture time for the highest algicidal activity were $30^{\circ}C$, pH 7 and above 24 hr, respectively. The algicidal activity were significantly increased at the stationary phase of the cell growth and continuously increased up to maximum at the death phase, probably due to the release of algicidal substance by cell Iysis. The effect of zinc ion addition on algicidal activity, was observed and indicated that the substance requires zinc for its activity. The candidate for the algicidal substance may be $\alpha$-mannosidase.

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Bacteriological Control of Cyanobacterial Bloom. (시안세균 수화의 세균학적 조절)

  • 김철호;권오섭;이진애
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.115-120
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    • 2004
  • A Gram (-), rod-shaped bacterium in size of 1.3∼$1.8{\times}0.35{\mu}m$ inhibiting the growth of cyanobacterium (Ana-baena cylindrica) was isolated and designated NG-2 in this manuscript. This isolate showed positive reactions for catalase and oxidase, and optimal growth conditions of 35∼TEX>$40<^{\circ}C$ and pH 9.0. In a mixed-culture of A. cylindrica and the isolate, each microorganism grew inverse-proportionally, and the cyanobacterial vegetative cells almost completely disappeared within 24 hours. NG-2 lysed A. cylindrica only under light, which means that lytic activity of NG-2 was dependent on the photosynthetic activity of host. When observed under phase contrast microscope, the isolate lysed vegetative cells of A. cylindrica in scattered state in a liquid medium, whereas het-erocysts have not been lysed. When cyanobacterial cell walls have been lysed partly, NG-2 attatched around A. cylindrica filament and formed colony, then encouraged complete lysis of cyanobacterial cells. The isolate showed similar lytic activity in natural water as in an artificial medium. And lytic activity of NG-2 was enhanced when attached on expandable polystyrene bead.

Isolation and Characterization of Alga-Lytic Bacterium HY0210-AK1 and Its Degradability of Anabaena cylindrica (남조류 분해세균 HY0210-AK1의 분리와 특성 및 Anabaena cylindrica 분해 활성)

  • 장은희;김정동;한명수
    • Korean Journal of Environmental Biology
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    • v.21 no.2
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    • pp.194-202
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    • 2003
  • To isolate alga-lytic bacteria, a number of samples were collected from Lake of Sukchon and Pal'tang reservoir where cyanobacteria blooming occurred. HY0210-AK1, which exhibited high alga-lytic activity, was isolated using Anabaena cylindrica lawn. The morphological and biochemical characteristics of the isolate HY0210-AK1 were very similar to that of the genus Rhizobium. Taxonomic identification including 16S rDNA base sequencing and phylogenetic analysis indicated that the isolate Hy0210-AK1 had a 99.1% homology in its 16S rDNA babe sequence with Sphingobium herbicidovorans. A. cylindrica NIES-19 was susceptible to the alga-lytic bacterial attack. The growth-inhibiting offset of the bacterium was not different on A. cylindrica NIES-19 when Sphingobium herbicidovorans HY0210-AK1 was in the lag, exponential, and stationary growth phase, although the alga-Iytic effect of S. herbici-dovorans HY0210-AK1 that in stationary growth phase was somewhat pronounced at the first time of inoculation. When S. herbicidovorans HY0210-AK1 was inoculated was inoculated with $1\times 10^{8}$ CFU $ml^{-1}$ together with A cylindrica NIES-19, the bacterium proliferated and caused algal lysis. A. cylindrica NIES-19 died when S. herbicidovorans HY0210 AKl was added to the algal culture but not when duly the filtrates from the bacterial culture was added. This suggests that extracellular substances are not responsible for inhibition of A. cylindrica NIES-19 and that algal Iysis largely attributed to direct interaction between S. herbicidovorans HY0210-AK1 and A. cylindrica NIES-19. The alga-lytic bacterium HY0210-AK1 caused cell lysis and death of three strain of Micro-cystis aeruginosa, but revealed no alga-Iytic effects on the Stephanodiscus hantzschii.