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Multi-drug Resistance of Mycoplasma pneumoniae Isolates from Patients with Respiratory Diseases against Quinolone and Macrolide (호흡기질환 환자에서 분리한 Mycoplasma pneumoniae의 Quinolone계와 Macrolide계 항생물질에 대한 다제 저항성)

  • Jun, Sung-Gon;Chang, Myung-Woong
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.435-443
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    • 2007
  • Antimicrobial susceptibility test of the 116 strains of Mycoplasma pneumoniae isolates were performed by a broth micro-dilution method against to moxifloxacin, levofloxacin, sparfloxacin, ofloxacin, ciprofloxacin, clarithromycin minocycline, erythromycin, josamycin, and tetracycline. The initial-minimum inhibitory concentration (I-MIC) was evaluated as the lowest concentration of antimicrobial agents that prevented a color change in the medium at that time when the drug-free growth control, about 7 days after incubation, and the final-minimum inhibitory concentration (F-MIC) was defined a color change about 14 days after incubation. The evaluation to the drug-resistant M. pneumoniae isolates were determined the $MIC{\pm}1.0$ ${\mu}g/ml$ of each antimicrobial agent. According to the I-MIC, single drug-resistant M. pneumoniae strains to ciprofloxacin, ofloxacin, clarithromycin and erythromycin were 79.3, 53.5, 10.3, and 7.8%, respectively. Two kinds of drug-resistant M. pneumoniae strains to ofloxacin and ciprofloxacin, or ciprofloxacin and clarithromycin were 42.2 and 9.5%. Three kinds of drug-resistant M. pneumoniae strains to erythromycin, ofloxacin, and ciprofloxacin, or ofloxacin, ciprofloxacin and clarithromycin were 6.9 and 6.0% . According to the F-MIC, single drug-resistant M. pneumoniae strains to tetracycline, ciprofloxacin, ofloxacin, minocycline,erythromycin, josamycin, clarithromycin and sparfloxacin were 91.4, 91.4, 91.4, 89.7, 68.1, 52.6, 28.5, and 11.2%, respectively. The incidence of two kinds of drug-resistant M. pneumoniae strains were from 20.7% to 91.4%, three kinds of drug-resistant M. pneumoniae strains were from 28.5% to 89.7%, four kinds of drug-resistant M. pneumoniae strains were 2.6%, five kinds of drug-resistant M. pneumoniae were from 2.6% to 21.6%, six kinds of drug-resistant M. pneumoniae strains were from 0.9% to 24.1%, seven kinds of drug-resistant M. pneumoniae strains were from 0.9% to 2.6%, and eight kinds of drug-resistant M. pneumoniae strains were 1.7%. These results suggest that sparfloxacin, moxifloxacin and levofloxacin might be promising antimicrobial agents for the treatment of M. pneumoniae infection in Korea. However, most strains of M. pneumoniae isolates were single or multi-resistance pattern to the other tested antimicrobial agents. Therefore, tetracycline, minocycline, erythromycin, clarithromycin, and second-generation quinolones are more carefully used to patients with M. pneumoniae infection in Korea.

Characterization of HtrA2-deficient Mouse Embryonic Fibroblast Cells Based on Morphology and Analysis of their Sensitivity in Response to Cell Death Stimuli. (HtrA2 유전자가 결손된 mouse embryonic fibroblast 세포주의 형태학적 특징 및 세포사멸 자극에 대한 감수성 조사)

  • Lee, Sang-Kyu;Nam, Min-Kyung;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.18 no.4
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    • pp.522-529
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    • 2008
  • High-temperature requirement A2(HtrA2) has been known as a human homologue of bacterial HtrA that has a molecular chaperone function. HtrA2 is mitochondrial serine protease that plays a significant role in regulating the apoptosis; however, the physiological function of HtrA2 still remains elusive. To establish experimental system for the investigation of new insights into the function of HtrA2 in mammalian cells, we first obtained $HtrA2^{+/+}$ and $HtrA2^{-/-}$ MEF cells lines and identified those cells based on the expression pattern and subcellular localization of HtrA2, using immunoblot and biochemical assays. Additionally, we observed that the morphological characteristics of $HtrA2^{-/-}$ MEF cells are different form those of $HtrA2^{+/+}$ MEF cells, showing a rounded shape instead of a typical fibroblast-like shape. Growth rate of $HtrA2^{-/-}$ MEF cells was also 1.4-fold higher than that of $HtrA2^{+/+}$ MEF cells at 36 hours. Furthermore, we verified both MEF cell lines induced caspsase-dependent cell death in response to apoptotic stimuli such as heat shock, staurosporine, and rotenone. The relationship between HtrA2 and heat shock-induced cell death is the first demonstration of the research field of HtrA2. Our study suggests that those MEF cell lines are suitable reagents to further investigate the molecular mechanism by which HtrA2 regulates the balance between cell death and survival.

Presence of Leukemia-maintaining Cells in Differentiation-resistant Fraction of K562 Chronic Myelogenous Leukemia (만성 골수성 백혈병 K562세포의 분화 내성 분획에서 백혈병 유지 세포의 동정)

  • Lee, Hong-Rae;Kim, Mi-Ju;Ha, Gahee;Kim, So-Jung;Kim, Sun-Hee;Kang, Chi-Dug
    • Journal of Life Science
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    • v.23 no.2
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    • pp.197-206
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    • 2013
  • The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.

Relationship Between Dinoflagellate Cyst Distribution in Surface Sediments and Phytoplankton Assemblages from Gwangyang Bay, a Southern Coastal area of Korea (한국 남해 연안 광양만 표층 퇴적물의 와편모조류 시스트 분포 특성과 식물플랑크톤 군집과의 비교)

  • 김소영;문창호;조현진
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.8 no.2
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    • pp.111-120
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    • 2003
  • To describe dinoflagellate cysts from Gwangyang Bay, surface sediment samples were collected at 20 sites by the TFO core sampler on 24 August 2001, in coupled with a phytoplankton investigation by surface seawater sampling. More than 17 genera, 36 species of dinoflagellate cysts were Identified from the sediment samples of Gwangyang Bay, consisting of 14 species of gonyaulacoid, 14 species of protoperidinioid, 3 species of diplopsalid, 2 species of gymnodinioid, 1 species of tuberculodinioid and calciodinellid, respectively. Cyst concentrations in Gwangyang Bay varied from 115 to 2,188 cysts/g, and generally increased toward a western part of the study area. The highest cyst concentration was observed at St. 11 located in the northwestern region with 11 genera and 19 species(2,188 cysts/g), while the lowest value with 6 genera and 9 species(115 cysts/g) was observed at St. 3 located in the center of the study area. The predominant dinoflagellate cyst was Spiniferites bulloideus, followed by Alexandrium sp., Brigantedinium simplex and S. delicatus. The motile forms of eight dinoflagellate cysts recorded in the sediment samples were also observed in the seawater: Polykrikos swartzii/kofoidii complex, Scripssiella trochoidea, Protoperidinium claudicans(cyst name: Votadinium spinosum), P. pentagonum(: Trinovantedinium capitatum capitatum), P. conicum(: Selenopemphix quanta), P. leonis(: Quinquecuspis concretum), P. conicoides(: Brigantedinium simplex), Gonyaulax spp.(: Spiniferites spp.). In this study, heterotrophic dinoflagellate cysts show the highest concentration at St. 6 where the highest density of diatoms simultaneously observed from surface water sample. This result suggests that the grazing of heterotrophic dinoflagellates on the diatoms in high concentration caused the higher concentration of heterotrophic dinoflagellate cysts.

Sedimentary facies and micropaleontological study of tidal sediments off the Mankyung-Dongjin River estuary, west coast of Korea. (한국 서해 만경강-동진강 하구역 및 연안역 조간대 퇴적층의 퇴적상과 미고생물학 적 연구)

  • 이영길;박용안
    • 한국해양학회지
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    • v.30 no.2
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    • pp.77-90
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    • 1995
  • The sedimentary environments and biostratigrapy of the tidal sediments off the Mankyung-Dongjin River estuary were studied based on sedimentary facies and diatom assemblage analysis. Sediment facies from the five vibracores are mainly clay, silt, and sand facies. The clay and silty sediment facies are more dominant than the sandy facies, and contain diatom frustules. The frequency of the diatom frustules are rate to common, but not found in sandy sediment facies. Bigeneric structures such as burrow and non-bigeneric primary sedimentary structures such as laminated sand and mud or silt and mud couplets, flaser bedding, ripple-cross lamination are found in several stratigraphic levels of the sedimentary sequences. A total of 219 species and varieties, belonging to 61 genera has been identified in the present study. Among them, paralia sulcata is the most abundant species about 30 to 50% of the total diatom frustules. Another predominant species are Cyclotella striata. Thalassionema nitzschioides. Actinoptychus undulatus. Delphineis surirella, Raphoneis amphiceros. Most of the diatoms occurred in this study area are marine, marine-brackish water, and brackish water species, and are benthic and tychopelagic or meroplanktonic species. Also, most of the species are coastal to littoral and littoral to inner neritic species. The occurrences of freshwater species, about 1 to 5% is higher than that of the Namyang Bay tidal sediments. The ecological properties of the diatoms occurred in the study area and primary sedimentary structure such as flaser bedding ripple cross bedding indicate that the deposits are formed under coastal or littoral to subeditorial environments such as tidal zone which was subjected to the influenced of stream water and was more strongly influenced by temperate to warm water than cold water. The sedimentary environments have not been changed distinctively during the time of deposition. The vertical distribution pattern of diatoms in the study area is studied by Q-mode cluster Analysis using spss/pc+ (ver.4.0). The results show that the three cores (GE-3, GE-11, GE-12) are divided into two diatom assemblages, respectively. Biostratigraphic correlation using the data of Q-mode Cluster Analysis are attempt in this study.

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Temporal and Spatial Variability of Phytoplankton Communities in the Nakdong River Estuary and Coastal Area, 2011-2012 (2011-2012년 낙동강 하구 및 연안역에서 식물플랑크톤 군집의 시·공간적 변화)

  • Chung, Mi Hee;Youn, Seok-Hyun
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.18 no.4
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    • pp.214-226
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    • 2013
  • To understand the changing patterns in phytoplankton communities, we conducted 12 surveys along the Nakdong River, its estuary, and adjacent coastal areas between January 2011 and October 2012 (during the period of barrage construction and sediment dredging). Monthly precipitation ranged from 0 to 502 mm during the survey period, and salinity ranged between 0.1 psu and 0.3 psu in the Nakdong River, regardless of the depth, indicating no seawater influence, while salinity showed large seasonal fluctuations in the estuarine and coastal station, ranging from 0.1 psu to 34.8 psu. A total of 402 phytoplankton species were identified, 178 species from the river and 331 species from the estuary and coastal areas. Phytoplankton standing crop increased in 2012 compared to that in 2011, and was found to be highest in the river, followed by the estuary and coastal areas. Among the top 20 species in frequency of occurrence and dominance, Stephanodiscus spp., Aulacoseira granulata, and Aulacoseira granulata var. angustissima and Pseudo-nitzschia spp. were important species along the river-estuary-coastal areas. Diatoms were the major taxonomic group inhabiting the Nakdong river-estuary-coastal areas. A comparison of seasonal dominant phytoplankton species revealed a slight decrease over the years, from 13 species in 2011 to 10 species in 2012. However, no significant difference was found in the diversity of phytoplankton species between the two survey years, although lightly greater diversity was observed in the coastal areas than in the river and estuary. Cluster analysis with community composition data revealed that the community structure varied significantly in 2011 depending on the time of survey, while in 2012, it hardly showed any variation and was simpler. An increase in the phytoplankton standing crop, fewer dominant species, and simpler community structure in 2012 compared to those in 2011 are probably due to the rapid environmental changes along the Nakdong River. To investigate these ecological relationships, it is necessary to conduct further studies focusing on integrated analyses of biocenosis, including phytoplankton with respect to the changes in nutrient distribution, variation of freshwater discharge, and effect area of freshwater in the Nakdong estuary and adjacent coastal areas.

Development of EvaGreen Based Real-time PCR Assay for Detection and Quantification Toxic Dinoflagellate Pfiesteria Piscicida and Field Applications (유독 와편모조류 Pfiesteria Piscicida 탐지 및 정량 분석을 위한 EvaGreen 기반 Real-time PCR기법 개발과 현장 적용)

  • PARK, BUM SOO;JOO, JAE-HYOUNG;KIM, MYO-KYUNG;KIM, JOO-HWAN;KIM, JIN HO;BAEK, SEUNG HO;HAN, MYUNG-SOO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.22 no.1
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    • pp.31-44
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    • 2017
  • Pfiesteria piscicida is one of heterotrophic dinoflagellate having toxic metaboliges, and it is difficult to detect and quantify this dinoflagellate via light microscope due to small size and morphological similarity with Pfiesteria-like dinoflagellate (PLD) species. Alternatively, we developed quantitative real-time PCR assay based on EvaGreen and determined field accessibility throughout the investigation of distribution in the entire Korean coastal waters and population dynamics in Shihwa Lake. The P. piscicida-specific primers based on internal transcribed spacer 1 (ITS 1) were designed and the specificity of primers was confirmed by PCR with other genomic DNAs which have genetic similarity with target species. Through real-time PCR assay, a standard curve which had a significant linear correlation between log cell number and $C_T$ value ($r^2{\geq}0.999$) and one informative melting peak ($88^{\circ}C$) were obtained. These results implies that developed real-time PCR can accurately detect and quantify P. piscicida. Throughout the field applications of real-time PCR assay, P. piscicida was distributed in western (Mokpo and Kimje) and easthern (Gangneng) Korean coastal water even though light microscopy failed to identify P. piscicida. In the investigation of population dynamics in Shihwa Lake, the density of P. piscicida was peaked in June, July and August 2007 at St. 1 where salinity (${\leq}15psu$) was lower than the other 2 sites. In this study, we successed to develop EvaGreen bassed real-time PCR for detection and quantification of P. piscicida in fields, so this developed assay will be useful for various ecological studies in the future.

Characterization of a Novel Fibrinolytic Enzyme Produced from Bacillus subtilis BK-17 (Bacillus subtilis BK-17 유래 혈전용해 효소의 특성)

  • Hyun Bek;Lim Hak-Seob;Chung Kyung Kae;Choi Yung Hyun;Choi Byung Tae;Seo Min-Jeong;Kim Ji-Eun;Ryu Eun-Ju;Huh Man Kyu;Joo Woo Hong;Jeong Young Kee
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.987-993
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    • 2005
  • A bacterium, producing a fibrinolytic enzyme, was screened from a decaying rice plant. The bacterium was identified as Bacillus subtilis by morphological, biochemical, and physiological properties and named Bacillus subtilis BK-17. The fibrinolytic enzyme (BK) was purified from supernatant of Bacillus subtilis BK-17 culture broth. The molecular weight was 31 kDa as determined by SDS-PAGE. The effect of temperature, pH, and plasminogen on the activity of the bacillokinase (BK) was analysed and the activity was compared with urokinase. The optimal temperature and pH were $50^{circ}C$ and pH 7, pH 8, respectively. The BK activity was inhibited to $45\%$, $35\%$, and $23\%$ with 1mM EDTA, $Zn^{2+}$, and $Ca^{2+}$, respectively. However, $Mg^{2+}$, $Mn^{2+}$, and $Co^{2+}$ ions did not have any significant effect on the enzyme activity The BK showed the artivity in the both plates, plasminogen-free fibrin plate and plasminogen-rich fibrin plate. The result indicates that the BK can directly act the fibrin. In comparison of fibrinolytic activity with urokinase on the fibrin plate, the BK shows about 20 folds higher activity than that of the urokinase.

Effect of the Hexane Extract of Saussurea lappa on the Growth of HT-29 Human Colon Cancer Cells (목향 헥산추출물이 대장암세포인 HT-29 세포의 증식에 미치는 영향)

  • Kim, Eun-Ji;Park, Hee-Sook;Lim, Soon-Sung;Kim, Jong-Sang;Shin, Hyun-Kyung;Yoon, Jung-Han
    • Korean Journal of Food Science and Technology
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    • v.40 no.2
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    • pp.207-214
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    • 2008
  • In Asia Saussurea lappa (SL) has been used as a traditional herbal medicine to treat abdominal pain and tenesmus. Recently, in vitro cell culture studies have shown that SL has anti-ulcer, anti-inflammatory, and anti-tumor properties. To explore its potential chemopreventive and chemotherapeutic effects in colon cancer, we examined whether the hexane extract of SL (HESL) could inhibit the growth of HT-29 human colon cancer cells, and investigated the mechanisms for this effect. The cells were cultured with various concentrations (0-5 ${\mu}g/mL$) of HESL. The results indicated that HESL markedly decreased the numbers of viable HT-29 cells; whereas at the concentration of 5 ${\mu}g/mL$, HESL slightly decreased the viable cell numbers of CCD 1108Sk human skin normal fibroblasts at 72 hr. HESL substantially increased the numbers of cells in the sub G1 phase, and dose-dependently increased apoptotic cell numbers. Western blot analysis of the total cell lysates revealed that HESL increased Bax protein levels, but did not affect Bcl-2 levels. HESL induced the cleavage of poly (ADP-ribose) polymerase and caspases 8, 9, 7, and 3. This study demonstrated that HESL inhibits cell growth and induces apoptosis in HT-29 cells, which may be mediated by its ability to increase Bax levels and activate the caspase pathway. These findings may lead to the development of new therapeutic strategies for colon cancer treatment.

Quality characteristics of distilled spirits by different nuruk-derived yeast (누룩 유래 효모의 종류에 따른 증류주의 품질 특성)

  • Lee, Ae-Ran;Kang, Sun-Hee;Kim, Hye-Ryun;Lee, Jang-Eun;Lee, Eun-Jung;Kim, Tae-Wan
    • Korean Journal of Food Science and Technology
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    • v.49 no.4
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    • pp.383-389
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    • 2017
  • This study aimed to select a yeast strain for optimizing the quality of distilled spirits. The brewing and distilling properties of 4 KFRI (Korea Food Research Institute) yeasts (Y88-4, Y98-4, Y172-6, Y192-4) and 2 industry yeasts (C1, C2) were compared. For investigating the possibility of using these strains on an industrial scale, diverse analytical methods were applied to assess parameters associated with distilled spirit quality such as alcohol content, pH, total acidity, and soluble solid content. After 11 days of fermentation, the alcohol strength obtained using six yeast strains reached 13.9-16.4% (v/v), while pH was 3.9-4.0, and total acid was 0.40-0.52%. To compare GC-MSD Volatile flavor components, all the distilled spirit samples were diluted to 20% (v/v) alcohol strength. Seven fusel alcohols, 26 esters, 2 acids, and 3 miscellaneous compounds were detected in the distilled spirits. Y88-4 had the most abundant volatile flavor component and scored the highest overall preference in sensory evaluation. After analyzing the various properties of yeasts, strain Y88-4 was finally selected as the best strain for producing distilled spirits.