• Title/Summary/Keyword: long-term culturing

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Laboratory Culture Media-Dependent Biocontrol Ability of Burkholderia gladioli strain B543

  • Bae, Yeoung-Seuk;Park, Kyung-Seok;Choi, Ok-Hee
    • The Plant Pathology Journal
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    • v.23 no.3
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    • pp.161-165
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    • 2007
  • Cultivation of a biocontrol agent on a certain medium often results in reduced biocontrol efficacy and alters physiological state. In our previous study, Burkholderia gladioli strain B543 with long-term subculture on tryptic soy agar resulted in significantly reduced biocontrol ability against cucumber damping-off caused by P. ultimum. Therefore, we investigated the influence of laboratory culturing media on biocontrol activity and physiological state of Burkholderia gladioli strain B543 by using long-term repeated culture on a certain medium. When isolate B543 were successionally cultured on King's B agar (KBA), tryptic soy agar, nutrient agar (NA), or soil extract agar more than 20 times, the isolate cultured on KBA or NA showed a significantly enhanced biocontrol efficacy and higher population density in the rhizosphere of cucumber compared to that of the others. However, the isolates cultured on KBA more than 20 times showed the lowest production of protease, siderophore, or antifungal substance(s), measured by skim milk agar, Chrome-Azurol-S agar, and potato dextrose agar amended with 10% of the culture filtrate, respectively. Our results suggest that adaptation to proper culturing medium can alter biocontrol ability and physiological state, and we must consider laboratory media in optimizing the use of biocontrol agents.

Effect of culturing media on biocontrol ability and physiological state of Burkholderia gladioli strain B543.

  • Bae, Yeoung-Seuk;Park, Kyungseok;Kim, Choong-Hoe
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.87.2-87
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    • 2003
  • Long-term repeated culturing of biocontrol agents on a certain medium often results in reduced biocontrol efficacy and altered physiology. Effect of culturing media on biocontrol ability and physiological state of Burkholderia gladioli strain B543 was investigated. Over 20 times repeated cultivation of B. giadioli strain B543 on Kings B medium or nutrient agar medium showed improved biological control of cucumber damping-off caused by Pythium ultimum, while one time cultivation on KB or NA did not. The repeated cultivation also induced the physiological changes of the biocontrol agent such as antifungal activity and the production of protease and siderophore. Our result indicates that adaptation to proper culturing medium can alter biocontrol ability and must consider in optimizing the use of biocontrol agents.

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Long-term Change and Factors Affecting the Fatness of the Pacific Oyster Crassostrea gigas in Tongyeong-Geoje Bays, Korea (통영-거제해역 수하연 양식 참굴(Crassostrea gigas)의 비만도 장기변화와 영향 요인 고찰)

  • Shim, JeongHee;Lee, Sang Jun;Koo, Jun-Ho;Jeong, Rae Hong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.4
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    • pp.434-444
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    • 2021
  • The decrease in fatness of the Pacific oyster Crassostrea gigas, which consequently results in decrease in the profit of aquaculture industry, has become a source of serious concern in southeast coast of Korea. The ratio of flesh (edible portion) to total oyster weight, commonly called edible portion yield ("suyul" in Korean), have been used as a fatness index for the healthy and valuable state of oyster from the early stage of oyster farming in Korea. More than 360 data sets were collected from early culturing periods (in the 1970s) to the present from approximately 15 published literatures to evaluate the long-term fatness trend of oyster, reared particularly in submerged longline culturing system in Gyeongsangnam-do province. Slight decrease in oyster fatness during the 1970s to 1990s was detected in Tongyeong and Geoje Bays; however, from the 1990s to the present, clear decreasing trends were observed with a decrease of 0.04-0.08% year-1, especially during harvest season. Oyster mass production per unit area almost doubled within a short period in the early 2000s; however, changes in coastal environment factors inhibited the fattening of shellfish from the mid 1990s. These results indicate that the severe competition in feeding and low biological production in water column might be some convincing reasons for the decrease in fatness of oyster from the 1990s, in Tongyeong and Geoje Bays, Korea.

Minoxidil Regulates Aging-Like Phenotypes in Rat Cortical Astrocytes In Vitro

  • Minji Bang;Seung Jin Yang;TaeJin Ahn;Seol-Heui Han;Chan Young Shin;Kyoung Ja Kwon
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.116-126
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    • 2023
  • Mainly due to the slanted focus on the mechanism and regulation of neuronal aging, research on astrocyte aging and its modulation during brain aging is scarce. In this study, we established aged astrocyte culture model by long-term culturing. Cellular senescence was confirmed through SA-β-gal staining as well as through the examination of morphological, molecular, and functional markers. RNA sequencing and functional analysis of astrocytes were performed to further investigate the detailed characteristics of the aged astrocyte model. Along with aged phenotypes, decreased astrocytic proliferation, migration, mitochondrial energetic function and support for neuronal survival and differentiation has been observed in aged astrocytes. In addition, increased expression of cytokines and chemokine-related factors including plasminogen activator inhibitor -1 (PAI-1) was observed in aged astrocytes. Using the RNA sequencing results, we searched potential drugs that can normalize the dysregulated gene expression pattern observed in long-term cultured aged astrocytes. Among several candidates, minoxidil, a pyrimidine-derived anti-hypertensive and anti-pattern hair loss drug, normalized the increased number of SA-β-gal positive cells and nuclear size in aged astrocytes. In addition, minoxidil restored up-regulated activity of PAI-1 and increased mitochondrial superoxide production in aged astrocytes. We concluded that long term culture of astrocytes can be used as a reliable model for the study of astrocyte senescence and minoxidil can be a plausible candidate for the regulation of brain aging.

Effect of Acid Rain in Soil Microorganism (산성비가 토양미생물에 미치는 영향)

  • Kim, Gab-Jung;Lim, Jin-A;Park, SeongJoo;Mun, Hyeong-Tae;Park, Kyeong-Ryang;Lee, In-Soo
    • Journal of Life Science
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    • v.8 no.3
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    • pp.299-304
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    • 1998
  • In order to clarify the effects of acid rain on soil microorganisms, the inpact of acid to soil microorganisms was survyed for 14 weeks using soil microcosms from industrial site A and B, Gaejok mountain, and Daechong lake in Taejeon area. The acid tolerant-microorganisms in natural soil, using culturing method were counted to be 5.8 - $8.0{\times}10^6$CFU/g soil. The number of microorganisms using ATP-biomass analysis for natural soil samples were also analyzed and 2.2 - $2.6{\times}10^9$ cell/g soil in industrial site A and B, Gaejok mountain, and Daechong lake were determined. In soil samples, which were treated with artificial acid rain, the number of acid tolerant microorganisms were counted 2.9 - $5.8{\times}10^5$ and 2.8 - $7.5{\times}10^8$, respectively. Therefore, we conformed that the numver of soil microorganisms were influenced by acid rain. Also, long term acid tolerant microorganisms were identified as Rhodotorula sp. and Pseudomonas sp.

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Nitrogen removal performance of anammox process with PVA-SA gel bead crosslinked with sodium sulfate as a biomass carrier

  • Tuyen, N.V.;Ryu, J.H.;Yae, J.B.;Kim, H.G.;Hong, S.W.;Ahn, D.H.
    • Journal of Industrial and Engineering Chemistry
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    • v.67
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    • pp.326-332
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    • 2018
  • In this study,the result shows that polyvinyl alcohol-sodium alginate (PVA-SA) gel bead crosslinked with sodium sulfate are better among the different methods by comparing the relative mechanical strength, mechanical strength swelling and expansion coefficient of beads in water. Subsequently, anammox biomass entrapment by PVA-SA gel was introduced into continuous stirred tank reactor (CSTR). After 24 operation days, the nitrogen removal efficiency achieved 60%, while the nitrogen loading rate (NLR) was $0.14kgN/m^3/d$ and the experiment data indicated that PVA-SA gel bead crosslinked with sodium sulfate can be used to initiate anammox process. Furthermore, it is an alternative for culturing anammox in a long-term operation.

Current Advances in Cryopreservation of Microalgae

  • Nugroho, Wahyu Sri Kunto;Kim, Do-A;Kim, Dong-Woo;Koo, Bon-Won;Hur, Young Baek;Kim, Hak Jun
    • Journal of Marine Life Science
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    • v.1 no.1
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    • pp.70-78
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    • 2016
  • Microalgae are of significant importance for future biotechnological applications. Many microalgae banks or laboratories attempt to maintain various microalgae for further research purposes. Cryopreservation has been preferred to reduce a labor-intensive and costly routine sub-culturing. Cryopreservation can also diminish the genetic drift risk. However, cryopreservation as a long term storage of microalgae method are still in developing progress because it cannot be generalized for all microalgae. Microalgae types, cryoprotectant agents (CPAs) types, freezing and thawing methods are the most important factors that should be considered for cryopreservation. In this short review the basic principles and the current advanced of microalgae cryopreservation methods are discussed with a suggested starting parameters for microalgae cryopreservation.

Application of Molecular Biology to Rumen Microbes -Review-

  • Kobayashi, Y.;Onodera, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.77-83
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    • 1999
  • Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

Continuous Stable production of won Willerand Factor Monoclonal Antibody in Spin Filter Bioreactor with Bleeding Technology

  • Yun, Joung-Won;Lee, Soo-Young;Park, Byung-Wook;Han-Kyu oh;Kim, Se-Ho;Byum, Tea-Ho;Park, Soung-yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.130-135
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    • 2000
  • The characteristics of two different modes of perfusion culture, intermittent and continuous bleedings, were investigated by culturing the hybridoma cells producing von Willebrand Factor (vWF) monoclonal antibody (McAb) in a 15 L bioreactor without clogging the filter. Both culture methods exhibited similar profiles of cell density and metabolite concentrations during the culture period at the cell concentration of around 1${\times}$107 cells/mL. When the perfusion rate was increased, the intermittrnt bleeding culture showed problems of ammonia accumulation and decrease of cell viability. The continuous bleeding culture in terms of nutrient consumption and metabolite production kinetics. But the analysis of specific oxygen consumption rate showed that the specific oxygen consumption rate of intermittent bleeding culture was similar to that of exponential growth phase. The continuous bleeding culture showed higher specific oxygen consumption rate of intermittent bleeding culture. finally we proved the possibility of long-term operation of continuous bleeding culture and produced approximately 40 g of vWF McAb in a 15L bioreactor after one-month operation.

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Diversity of Ion Channels in Human Bone Marrow Mesenchymal Stem Cells from Amyotrophic Lateral Sclerosis Patients

  • Park, Kyoung-Sun;Choi, Mi-Ran;Jung, Kyoung-Hwa;Kim, Seung-Hyun;Kim, Hyun-Young;Kim, Kyung-Suk;Cha, Eun-Jong;Kim, Yang-Mi;Chai, Young-Gyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.6
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    • pp.337-342
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    • 2008
  • Human bone marrow mesenchymal stem cells (hBM-MSCs) represent a potentially valuable cell type for clinical therapeutic applications. The present study was designed to evaluate the effect of long-term culturing (up to $10^{th}$ passages) of hBM-MSCs from eight individual amyotrophic lateral sclerosis (ALS) patients, focusing on functional ion channels. All hBM-MSCs contain several MSCs markers with no significant differences, whereas the distribution of functional ion channels was shown to be different between cells. Four types of $K^+$ currents, including noise-like $Ca^{+2}$-activated $K^+$ current ($IK_{Ca}$), a transient outward $K^+$ current ($I_{to}$), a delayed rectifier $K^+$ current ($IK_{DR}$), and an inward-rectifier $K^+$ current ($K_{ir}$) were heterogeneously present in these cells, and a TTX-sensitive $Na^+$ current ($I_{Na,TTX}$) was also recorded. In the RT-PCR analysis, Kv1.1,, heag1, Kv4.2, Kir2.1, MaxiK, and hNE-Na were detected. In particular, ($I_{Na,TTX}$) showed a significant passage-dependent increase. This is the first report showing that functional ion channel profiling depend on the cellular passage of hBM-MSCs.