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The Growth and EPA Synthesis of Shewanella oneidensis MR-1 and Expectation of EPA Biosynthetic Pathway

  • Jeong, Young-Su;Song, Sang-Kyu;Lee, Su-Jin;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.127-133
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    • 2006
  • Shewanella oneidensis MR-1 has the ability to inhale certain metals and chemical compounds and exhale these materials in an altered state; as a result, this microorganism has been widely applied in bioremediation protocols. However, the relevant characteristics of cell growth and biosynthesis of PuFAs have yet to be thoroughly investigated. Therefore, in this study, we have attempted to characterize the growth and fatty acid profiles of S. oneidensis MR-1 under a variety of temperature conditions. The fastest growth of S. oneidensis MR-1 was observed at $30^{\circ}C$, with a specific growth rate and doubling time of $0.6885h^{-1}\;and\;1.007 h$. The maximum cell mass of this microorganism was elicited at a temperature of $4^{\circ}C$. The eicosapentaenoic acid (EPA) synthesis of S. oneidensis MR-1 was evaluated under these different culture temperatures. S. oneidensis MR-1 was found not to synthesize EPA at temperatures in excess of $30^{\circ}C$, but was shown to synthesize EPA at temperatures below $30^{\circ}C$. The EPA content was found to increase with decreases in temperature. We then evaluated the EPA biosynthetic pathway, using a phylogenetic tree predicted on 16s rRNA sequences, and the homology of ORFs between S. oneidensis MR-1 and Shewanella putrefaciens SCRC-2738, which is known to harbor a polyketide synthase (PKS)-like module. The phylogenetic tree revealed that MR-1 was very closely related to both Moritella sp., which is known to synthesize DHA via a PKS-like pathway, and S. putrefaciens, which has been reported to synthesize EPA via an identical pathway. The homology between the PKS-like module of S. putrefaciens SCRC-2738 and the entire genome of S. oneidensis MR-1 was also analyzed, in order to mine the genes associated with the PKS-like pathway in S. oneidensis MR-1. A putative PKS-like module for EPA biosynthesis was verified by this analysis, and was also corroborated by the experimental finding that S. oneidensis MR-1 was able to synthesize EPA without the expression of $dihomo-{\gamma}-linoleic$ acid (DGLA) and arachidonic acid (AA) formed during EPA synthesis via the FAS pathway.

Collection of Soil Actinomycetes from Cheju Island and Screening for their Antibacterial Activities (제주도 토양방선균의 수집과 항균물질 생산균의 선별)

  • Chung, Wan-Seok;Kim, Chang-Jin;Ko, Young-Hwan
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.99-104
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    • 1999
  • Soil actinomycetes of 703 strains were isolated from 25 sampling points in Cheju Island using 4 different media. Arginine glycerol salts agar containing soil extract was found to be the best medium for the isolation of soil actinomycetes. Soil samples from pasture land showed higher population and diversity of the actinomycetes than those from citrus field, forest, island, hill or valley. When the antibacterial activity of the 526 isolates was tested against three bacterial strains, Escherichia coli, Staphylococcus aureus and Pseudomonas solanacearum the frequency of the isolates with antibacterial activity varied much depending upon the media used for isolation and cultivation. BL106Ba, one of the 10 isolates that showed antibacterial activity against all the above 3 test strains, was chosen based upon the pH and heat stability of its antibacterial metabolites, and was identified as Streptomyces sp. based upon its cultural, morphological and physiological characteristics. The partially purified white crystalline substance obtained from the culture supematant of BL1063a through cation exchange chromatography(AG MP-50) and three times consecutive gel filtration(Sephadex G-10) showed high antimicrobial activity against gram positive and negative bacteria, but low activity against yeasts. The partially purified substance was found to contain at least four different compounds with antibacterial activity by both thin layer chromatography and high performance liquid chromatography.

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Expression Analysis of the Ligand to Ly-6E.1 Mouse Hematopoietic Stem Cell Antigen

  • Hwang, Dae-Youn;Min, Dul-Lei;Sonn, Chung-Hee;Chang, Mi-Ra;Lee, Mi-Hyun;Paik, Sang-Gi;Kim, Young-Sang
    • Animal cells and systems
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    • v.1 no.1
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    • pp.157-164
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    • 1997
  • Ly-6E.1 antigen was proposed as a regulatory molecule of T lymphocyte activation, a hematopoietic stem cell marker, a memory cell marker, and an adhesion molecule. Though there were several reports suggesting the presence of Ly-6 ligand, the characterization of the ligand was not yet performed, As an attempt to screen the expression of Ly-6E.1 ligand, we prepared a probe for detecting Ly-6E.1 ligand by producing a fusion protein between Ly-6E.1 and $hlgC_{r1}$, A mammalian cell expression vector with Ly-6E.$1/hlgC_{r1}$ chimeric cDNA was transfected in SP2/0-Ag14 myeloma cells, and stable transfectants were selected. The fusion protein was produced as a dimer and maintained the epitopes for monoclonal antibodies specific for Ly-6E.1 and for anti-human lgG antibody. The purified fusion protein through Gammabind G column was used for FACS analyses for the expression of Ly-6E.1 ligand. The fusion protein interacted with several cell lines originating from B cells, T cells, or monocytes. The fusion Protein also strongly stained bone marrow, lymph node, and spleen cells, but thymic cells weakly, if any. The staining was more obvious in C57BL/6 $(Ly-6^b)$ than Balb/c $(Ly-6^a)$ mice. These results suggest that the interaction of Ly-6E.1 with Ly-6E.1 ligand may function both in the stem cell environment and in the activation of mature lymphocytes. The fusion protein may be a valuable tool in characterization of biochemical properties of the Ly-6E.1 ligand and, further, in isolating its cDNA.

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Fermentation of Cucurbita maxima Extracts with Microganisms from Kimchi (김치 유래 유산균을 이용한 단호박 발효음료 제조 기술 개발)

  • Roh, Hyun-Ji;Kim, Gi-Eun
    • KSBB Journal
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    • v.24 no.2
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    • pp.149-155
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    • 2009
  • 19 strains, which could be identified as Lactobacillus sp. were isolated. The Cucurbita maxima has been known as a traditional healthy food and variable positive effects on the human body were already reported. In this study we tried to develop a production process for a healthy fermented drink with Cucurbita maxima and strains originated from Kimchi. Many kinds of lacctobacci species existed in the fermented food cannot survive in the acidic conditions in the stomach. So we tried to search and select a strain, which can arrive to the small intestine. A species of a Lactobacillus named as C332 was identifed as Lactobacillus plantarum and selected for the fermentation process. With the treatment with artificial gastric juice and artificial bile the survival rate of the cells could be calculated. The physiological characteristics at the variable conditions have been tested. After fermentation process the sensoric tests on the product with panels were tried. The most of the cells could survive in the acidic conditions and falcultive anaerobe. Especially some antibacterial effects aganinst E.coli were also found. With all kinds of the results from our research the fermented Cucurbita maxima drink can be a successful item in the market.

Evaluation of Color Reproduction Characteristics of TFT-LCD and AMOLED for Mobile Phone (모바일폰용 TFT-LCD와 AMOLED의 색재현성 평가)

  • Park, Tae-Yong;Lee, Cheol-Hee;Ha, Yeong-Ho
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.45 no.1
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    • pp.29-37
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    • 2008
  • With the rapid development of display technology as well as the diversity of display, image quality assessment plays an important role in display color reproduction. The image quality has described by objective evaluation based on the physical measured data, such as contrast ratio, color reproduction range, color temperature, and gamma characteristics, etc., however, the final judgement is achieved by perceptual quality assessment of observers. Therefore, it is necessary to investigate the relationship between physical performance of display and perceptual image quality. Thus, in order to evaluate objectively the color reproduction characteristics of two mobile displays, TFT-LCD and AMOLED, we measured the darkroom contrast ratio of full screen, luminance and color temperature of full-screen white, full-screen gray and color linearity, tone reproduction of color scales for each RGB primary, and color reproduction range. Then the pair-wise comparison is implemented for image experts and naive observers not only to judge their preference on principal evaluation attributes but also to subjectively describe perceived image quality. Through two evaluation processes, we derived the objective bases that can prove the results of subjective image quality assessment by interconnecting physical factors that have influence on observers' preference judgements. Finally, this paper provided important information for improving the image quality and increasing preference from an observer's point of view.

Effect of Chitin Compost on Biological control of Fusarium wilt in Tomato Field (키틴퇴비를 이용한 토마토의 Fusarium 시들음병의 생물학적 제어)

  • Jin, Rong-De;Cho, Min-Young;Kim, Sung-Jae;Ryu, Ji-Yeon;Chae, Dong-Hyeon;Kim, Yong-Woong;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.1
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    • pp.15-20
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    • 2006
  • Biological control by chitinolytic microorganisms is being evaluated as management options for soilborne diseases. Forty kilograms of chitin compost (CTC) and control compost (CC) were amended on tomato plots ($15m{\times}0.5m$) 7 d before transplanting to evaluate enzymatic activities and the control of Fusarium wilt. Samples were taken on day 1, 3, 5, and 7, the day 1 corresponded to the 66 d after transplanting, the day on which the initial wilting symptoms occurred in plants of CC treated plots. The chitinase activity in soil of CTC was always higher compared to the control. Pathogenesis related (PR) protein (chitinase, ${\beta}$-1, 3-glucanase and peroxidase) activities in tomato roots in CC increased every day and showed marked differences compared to CTC. Wilting symptoms (96 d after transplanting) were reduced by 25% in CTC compared to the control. Protection of tomato plant may be correlated with the high levels of soil enzyme activities resulting from the chitin compost.

Interactions between common heterotrophic protists and the dinoflagellate Tripos furca: implication on the long duration of its red tides in the South Sea of Korea in 2020

  • Eom, Se Hee;Jeong, Hae Jin;Ok, Jin Hee;Park, Sang Ah;Kang, Hee Chang;You, Ji Hyun;Lee, Sung Yeon;Yoo, Yeong Du;Lim, An Suk;Lee, Moo Joon
    • ALGAE
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    • v.36 no.1
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    • pp.25-36
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    • 2021
  • The mixotrophic dinoflagellate Tripos furca causes red tides in the waters of many countries. To understand its population dynamics, mortality due to predation as well as growth rate should be assessed. Prior to the present study, the heterotrophic dinoflagellates Noctiluca scintillans, Polykrikos kofoidii, Protoperidinium steinii, and mixotrophic dinoflagellate Fragilidium subglobosum were known to ingest T. furca. However, if other common heterotrophic protists are able to feed on T. furca has not been tested. We explored interactions between T. furca and nine heterotrophic dinoflagellates and one naked ciliate. Furthermore, we investigated the abundance of T. furca and common heterotrophic protists in coastal-offshore waters off Yeosu, southern Korea, on Jul 31, 2020, during its red tide. Among the tested heterotrophic protists, the heterotrophic dinoflagellates Aduncodinium glandula, Luciella masanensis, and Pfiesteria piscicida were able to feed on T. furca. However, the heterotrophic dinoflagellates Gyrodiniellum shiwhaense, Gyrodinium dominans, Gyrodinium jinhaense, Gyrodinium moestrupii, Oblea rotunda, Oxyrrhis marina, and the naked ciliate Rimostrombidium sp. were unable to feed on it. However, T. furca did not support the growth of A. glandula, L. masanensis, or P. piscicida. Red tides dominated by T. furca prevailed in the South Sea of Korea from Jun 30 to Sep 5, 2020. The maximum abundance of heterotrophic dinoflagellates in the waters off Yeosu on Jul 31, 2020, was as low as 5.0 cells mL-1, and A. glandula, L. masanensis, and P. piscicida were not detected. Furthermore, the abundances of the known predators F. subglobosum, N. scintillans, P. kofoidii, and Protoperidinium spp. were very low or negligible. Therefore, no or low abundance of effective predators might be partially responsible for the long duration of the T. furca red tides in the South Sea of Korea in 2020.

Outbreak of Red Tides in the Coastal Waters off the Southern Saemankeum areas, Jeonbuk, Korea 1. Temporal and Spatial Variations in the Phytoplankton Community in the Summer-fall of 1999 (전북 새만금 남쪽 해역의 유해성 적조 발생연구 1. 1999년도 여름-가을 식물플랑크톤의 시공간적 변화)

  • 유영두;정해진;심재형;박재연;이경재;이원호;권효근;배세진;박종규
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.7 no.3
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    • pp.129-139
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    • 2002
  • We investigated the outbreak of red tides dominated by harmful dinoflagellates from August to November 1999 in the coastal waters off the southern Saemankeum areas where a huge red tide dominated by Cochlodinium polykrikoides had been first observed in 1998. We took water samples from 2~5 depths of 4 stations (with 3-4 additional stations during red tides) in this study period and then measured the abundance of phytoplankton, water temperature, salinity, and the concentrations of nutrients. In the study period harmful dinoflagellates Alexandrium tamarense, C. polykrikoides, Gymnodnium catenatum, Gyrodinium aureolum, Gymnodnium impudicum were present, and of these G. aureolum and C. polykrikoides formed red tide patches on September 16 and October 18, respectively. The date of the outbreak of red tide dominated by C. polykrikoides in the study area was approximately 50 days later than that off the Kohung areas in 1997 and the surface water temperature when the red tides outbroke in the former area was 6$^{\circ}C$ lower than that fur the latter area. The maximum abundance of C. polykrikoides on September 16, October 7 and 18 were 5, 14, and 463 cells $m\ell$$^{-1}$ , respectively. The growth rate of C. polykrikoides, isolated from the study area, was 0.3~0.4 d$^{-1}$ at 20~$25^{\circ}C$, which enable this species to reach the maximum concentration without being transported from the adjacent waters containing already made red tide patches. The outbreaks of red tides dominated by C. polykrikoides in the study area and off Kohung have occurred when and/or where the concentrations of diatoms were low. This evidence suggests that the outbreak of red tides dominated by C. polykrikoides is adversely affected by the high diatom concentrations or the conditions favorable for the growth of diatoms.

Transarterial embolization of renal artery in dogs with experimental hydronephrosis (개의 실험적 수신증에 실시한 신동맥 색전술)

  • Chang, Dongwoo
    • Korean Journal of Veterinary Research
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    • v.41 no.3
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    • pp.437-445
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    • 2001
  • This study was performed to validate the procedure of transarterial embolization of the renal artery (TAE-RA) using iohexol-ethanol solution in dogs with unilateral experimental hydronephrosis and to evaluate the embolized kidney using B-mode ultrasonography and selective angiography. Experimental hydronephrosis was induced by ligation of unilateral ureter in 12 dogs. Ultrasonographic findings revealed that size of the kidney was significantly increased at 9 days and 17 days and the length of renal cortex was significantly decreased at 17 days after ligation of the unilateral ureter and it was in accordance with dilation of ipsilateral renal pelvis. No significant change of BUN, creatinine, ALT, calcium, and phosphorus was found immediately after unilateral experimental hydronephrosis. Therefore, it was concluded that unilateral hydronephrosis was established in 12 dogs at 17 days after ligation of ureter. Renal artery embolization was performed using selective catheterization in the hydronephrotic kidney of seven dogs and EKG, $SpO_2$, body temperature, pulse, and respiratory rate were within normal ranges during procedures. Iohexol-ethanol solution was used as embolic material. Average ethanol dose for renal artery embolization was $1.94{\pm}1.24ml/kg$. There were no dogs expired after TAE-RA and no side effects associated with regurgitation of iohexol-ethanol solution. Revascularization of renal artery was not found in angiography in dogs treated by TAE-RA at immediately after TAE-RA and 14 days after TAE-RA. Ultrasonographically, the mean longitudinal length of the embolized kidney decreased significantly at 2 and 3 months after TAE-RA compared to that of contralateral normal kidney. In summary, marked shrinkage of the embolized kidney was observed in dogs with unilateral experimental hydronephrosis treated by TAE-RA with iohexol-ethanol and no adverse effects were observed throughout the observation period. It is concluded that TAE-RA with iohexol-ethanol solution is a viable alternative to nephrectomy in dogs with unilateral hydronephrosis.

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Toxicity of Persistent Organic Pollutants, PAHs and TBT, in Zooplankton and Influence on Their Viability (동물플랑크톤에 대한 지속성 유기오염물질 PAHs와 TBT의 독성 및 생존능력에 미치는 영향)

  • Jang, Poong-Guk;Shin, Kyung-Soon;Jang, Min-Chul;Park, Dong-Won;Jang, Man
    • Korean Journal of Environmental Biology
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    • v.22
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    • pp.1-10
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    • 2004
  • We conducted three experiments to estimate the toxicity of POPs (persistent organic pollutants) on two copepod species (Acartia erythraea and A. omorii) and Artemia sp.; (1) 48 h-LC$_{50}$ of A. omorii with the five PAHs [polycyelic aromatic hydrocarbons anthracene, benzo〔a〕pyrene, fluoranthene, phenanthrene, pyrene〕 which were often detected in the Gwangyang Bay, (2) toxicity of benzo〔a〕pyrene and TBT on Artemia in different temperatures (1$0^{\circ}C$, 15$^{\circ}C$, 2$0^{\circ}C$), (3) effects of benzo〔a〕pyrene and TBT on egg Production rate, hatching rate and fecal Pellet Production of two copepod species (A. erythraea and A. omorii) fed on Heterocapsa triquetra (dinoflagellate) exposed in benzo〔a〕pyrene. Toxic chemicals which were most effective to A. omorii were flueranthene (48 h-LC$_{50}$ 19.20 $\mu\textrm{g}$ L$^{-1}$ ) and benzo〔a〕pyrene (48 h-LC$_{50}$ 29.89 $\mu\textrm{g}$ L$^{-1}$ ). The toxi- city of chemicals to Artemia increased when temperature increased. The toxicity of TBT was about 100 times higher than that of benzo〔a〕pyrene at 15$^{\circ}C$. Food materials (Heterocapsa triquetra) exposed in benzo〔a〕pyrene, affected negatively the rate of egg production, hatching rate and the fecal pellet production of the copepods at the high concentration. It is suggested that an increase in the concentration of benzo〔a〕pyrene might offset the production of copepods in marine ecosystems. This study suggests that copepods may be used as n indicator for early warning of the risk of POPs in marine ecosystems.