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C-terminal Fusion of EGFP to Pneumolysin from Streptococcus pneumoniae modified its Hemolytic Activity (Streptococcus pneumoniae가 생산하는 pneumolysin의 EGFP 융합으로 인한 용혈활성 변화)

  • Chung, Kyung Tae;Lee, Jae Heon;Jo, Hye Ju
    • Journal of Life Science
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    • v.28 no.1
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    • pp.99-104
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    • 2018
  • Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.

Design of the Detector Head for Single Photon Detection in Breast Cancer Diagnosis and Its Performance Evaluation (유방암진단에서의 단일광자검출을 위한 검출기 전단부의 설계와 성능평가)

  • Kim, Kwang-Hyun;Cho, Gyu-Seong;Chung, Woon-Kwan
    • Journal of Radiation Protection and Research
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    • v.28 no.4
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    • pp.263-270
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    • 2003
  • Monte Carlo simulation has been peformed to induce optimized parameters of the detector head of gamma camera for the diagnosis of breast cancer and to evaluate it under the diagnosis condition of the breast cancer. For the simulation, we used Tungsten collimator, having a lattice structured array with holes of $3mm{\times}3mm$ and septal thickness of 0.25 mm, which are corresponding to the pixellated photosensor. For driving optimum parameters we used Trade-Offs procedure between the geometric efficiency and the spatial resolution, varying the detector head components. In order to pre-evaluate the performance of the optimized detector head, we assumed diagnosis condition that the breast tumor is located in the middle of phantom with various sizes and its location is 25 mm from the collimator surface, considering background count caused by radiation sources from other organs. It was shown that the performance of the optimized detector head can be degraded according to the breast cancer size and the background count under real diagnosis conditions of breast cancer. Therefore, it is concluded that the spatial resolution, which is used as an indicator to distinguish the various sizes of breast cancer and is dependent on the characteristic of the detector head, appears to be meaningless in early diagnosis of the breast cancer.

A Study on the High Pressure Pump Simulation Model of a Diesel Injection System (디젤 분사시스템의 고압펌프 시뮬레이션 모델에 대한 연구)

  • Kim, Joongbae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.10
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    • pp.102-109
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    • 2017
  • The high pressure pump of a diesel injection system compresses the fuel supplied at low pressure into high pressure fuel and maintains the fuel of the common rail at the required pressure level according to the engine operating conditions. The high pressure pump is required to operate normally in order to compress the fuel to a high pressure of 2000 bar during the entire lifetime of the vehicle. Consequently, a suitable design technique, material durability and high precision machining are required. In this study, the high pressure pump simulation model of a 1-plunger radial piston pump is modelled by using the AMESim code. The main simulation parameters are the displacement, flow rate and pressure characteristics of the inlet and outlet valves, cam torque characteristics, and operating characteristics of the fuel metering valve and overflow valve. In addition, the operating characteristics of the pump are simulated according to the parameter changes of the hole diameter and the spring initial force of the inlet valve. The simulation results show that the operation of the developed pump model is logically valid. This paper also proposes a simulation model that can be used for current pump design changes and new pump designs.

Film Properties of MOCVD TiN prepared by TDMAT and TDMAT/$NH_3$ (TDMAT와 TDMAT/$NH_3$ 로 형성한 MOCVD(Metal Organic Chemical Vapor Deposition) Titanium Nitride 박막의 특성)

  • Baek, Su-Hyeon;Kim, Jang-Su;Park, Sang-Uk;Won, Seok-Jun;Jang, Yeong-Hak;O, Jae-Eung;Lee, Hyeon-Deok;Lee, Sang-In;Choe, Jin-Seok
    • Korean Journal of Materials Research
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    • v.5 no.7
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    • pp.775-780
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    • 1995
  • Thin films of titanium nitride are formed using the tetrakis-dimethyl-amino-titanium (TDMAT(Ti[N($CH_3$)$_2$]$_4$)) under various conditions. The formation of TiN films has been obtained from the thermal decomposition of the Ti-precursor and the gas phase reaction between TDMAT and ammonia(NH$_3$). The resistivity of the MOCVD film can be attributed to their impurity. Especially the curve fitting graph of XPS data is revealed that main impurities in the films as carbon and oxygen make various interstitial compounds which has influenced physical and electrical properties of the film. In the contact hole with the aspect ratio of 3:1 and the diameter of 0.5${\mu}{\textrm}{m}$, the SEM morphology shows that the step coverage is more decreased in the films formed y flowing ammonia additionally than the films formed by pyrolysis of TDMAT and the phenomenon is probably related with the activation energy.

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Biochemical and Cytological Changes of Pseudomonas sp. DJ-12 Cells in Response to Catechol Treatment (Catechol 처리에 의한 Pseudomonas sp. DJ-12의 생화학 및 세포학적 변화)

  • 고연자;임재윤;김치경;이기성
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.139-145
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    • 1999
  • Aromatic hydrocarbons which are not easily degraded by microorganisms can be accumulated in the conlaminated environment for a long lime, producing toxic effects on wild lives and humans. However, the sublethal concentrations of the chemicals induce the synthesis of stress-shock proteins in the cells and increase the adaptability of the organisms to the environmental stresses. In this study, therefore, the cells of Psezido~nonus sp. DJ- 12 treated with catechol at various concentrations were inveshgated for their survival, biodegtadability of catechol, production of stress-shock proteins, and cytological changes. The organisms were capable of degrading catechol at the range of 0.5 to 1.0 mM concentration wilhin 6 hours incubation, but they were killed by $10^2$-10$^3$ celllinl at 3 mM or higel- concentration without any catechol degradation. These cells treated with catechol begm lo produce DnaK and GroEL at 1 mM and 0.5 mM. respectively. Pseudumonas sp. DJ-12 treated with 10 mM catechol for I hour exhihiled some punctuated pores on the cell wall and contortion of the rod shape. The cells treated with he sublethal concentration of catechol showed the increased tolerance for suvival when exposed to the lethal concentration, and such tolerant effects were functioned crossly among benzoate, 4-chlorobenzoate, 'and catechol.

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An Experimental Study on the Bolted Connection Fatigue Capacity of Corrugated Steel Plates (파형강판 볼트 이음부의 피로성능에 관한 실험적 연구)

  • Oh, Hong-Seob
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.18 no.2
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    • pp.54-63
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    • 2014
  • Corrugated steel plate structure, which is built by assembling corrugated steel plate segments with bolts on site and filling the surroundings with quality soil, is widely used for buried structures as a eco-corridors, small bridges, and closed conduits. This experimental study is dealt with the static and fatigue performance of bolt connected corrugated steel plates under flexural loading. The experimental variables to verify the fatigue performance are bolt diameters and detailing of connection such as washer and the corrugation dimension of specimens has a $400{\times}150$ mm. The experimental ultimate strength of specimens under static loading was higher than the theoretical strength and all specimen failed by a bearing and tearing failure of bolt hole of upper plate. Therefore, a fatigue tests of specimens had 6.0mm and 7.0mm thickness was conducted in which the load range was up to 209kN and 516kN, respectively. From the fatigue test, failure patterns are changed from plate bearing and tearing which is a typical failure pattern of static failure to a bearing failure of plate and shear failure of bolt, and experimental fatigue limit at $2{\times}10^6$cycles is about 85MPa.

Effect of Bean Water Concentration and Incubation Time on Amylase Activity and Physicochemical Characteristics of Yukwa Paste (콩물 농도 및 숙성 시간이 Amylase 의 활성과 유과 반죽의 특성에 미치는 영향)

  • Sohn, Kyung-Hee;Jo, Mi-Na;Jeon, Hyeong-Ju;Park, Jin;Joo, Myung-Sook
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.288-293
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    • 2001
  • ${\alpha}-amylase$ activities of bean water was not significantly influenced by bean water concentrations but they were remarkably influenced by different temperatures and substrates. ${\alpha}-amylase$ activities of bean water on cooked starch were significantly higher than those on raw starch. ${\beta}-amylase$ and glucoamylase activities in 14% bean water were significantly higher than those in 7% bean water. Yukwa paste is glutinous rice flour paste. Bean water was added to Yukwa paste by 0, 7, 14% and incubated 0, 3, 6, 9, 12 hours at $60^{\circ}C$. The pH of Yukwa paste increased with bean water concentration and decreased with the incubation time. The viscosity decreased with bean water concentration and incubation time. The ruducing sugar content of Yukwa paste increased with bean water concentration and incubation time. The changes of reducing sugar content in cooked Yukwa paste were much higher than those in the raw one. ${\alpha},\;{\beta}-amylase$glucoamylase activities of Yukwa paste also increased with bean water concentration, and their activities were much higher on the cooked glutinous rice flour than those on the raw one. The SEM observation on the freeze-dried flour of Yukwa paste showed breakdown of amylopectin structure by addition of bean water in the paste.

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Distribution and Collection of Osmia Bees in the Mountain Areas of Korea (산지에 서식하는 뿔가위벌류의 분포와 채집)

  • 이순원;최경희;이동혁;김동아;류하경;이영인
    • Korean journal of applied entomology
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    • v.41 no.4
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    • pp.263-267
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    • 2002
  • A faunistic survey was conducted to find Osmia bees to be used as an effective pollinator in apple orchards. Collections were carried out from more than 50 selected sites in mountainous areas throughout Korea for 3 years (1996-98) with providing bundle of bamboo traps (6-8 mm of inner diameter) as Osmia nesting site. Five species of Osmia :O. cornifrons (Radoszkowsky), O. pedicornis Cockerell, O. taurus Smith, O. satoi Yasumatsu et Hirashima and O. jacotti Cockerell, were collected at the rates of 39.9-51.7, 28.3-53.7, 5.5-19.1, 0.8-0.9 and 0.0-0.1%, respectively. And O. cornifrons and O. pedicornis were the dominants with over 80% of total Osmia collected through the collections. Out of those bees collected, rates of female were 29-43% for O. cornifrons. 10-12% for O. pedicornis and 22-43% for O. taurus. When the collection sites of the dominant 3 species were grouped in relation to altitudes above the sea level, about 50% or more bees were collected form the higher sites (500m or higher) while 20% or less were from the lower sites (200m or lower). When the bamboo traps were placed at the same sites continuously for 3 years, number of O. cornrfrons and O. taurus collected was slightly increased while that of O. pedicornis was drastically decreased year after year. Nesting by Osmia bees seems to have started at around early April, peaked at early May and ended by Mid June, while nest competing species of Sphecidae and Eumenidae started nesting at around early June, peaked at early July and ended by the Mid August.

Taxonomic study on Korean Aphyllophorales (II) -on some unrecorded species- (한국산 민주름버섯목의 분류학적 연구 (II) -수종 미기록종에 대하여-)

  • Jung, Hack-Sung
    • The Korean Journal of Mycology
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    • v.24 no.3 s.78
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    • pp.228-236
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    • 1996
  • Flesh fungi were collected during field trips to mountain areas throughout the country from May to October of 1994. Through the observation and identification of specimens belonging to the Aphyllophorales, one genus, Tylospora, and six species, Athelia fibulata, Hypochnicium punctulatum, Tylospora fibrillosa, Stereum ochraceo-flavum, Steccherinum litschaueri, and Oligoporus undosus were confirmed new to Korea and are registered here with descriptions.

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Anxiolytic Effects of Quercetin: Involvement of GABAergic System (Quercetin의 항불안 효과: GABA 신경계를 중심으로)

  • Jung, Ji Wook;Lee, Seungheon
    • Journal of Life Science
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    • v.24 no.3
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    • pp.290-296
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    • 2014
  • The present experiment investigated putative anxiolytic-like effects of quercetin using an elevated plus-maze (EPM) and hole-board apparatus test in mice. Quercetin is a flavonoid widely distributed in nature. Quercetin (1.25, 2.5, 5, or 10 mg/kg) was orally administered to ICR mice 1 h before a behavioral evaluation in the EPM. Control mice were treated with an equal volume of vehicle, and positive control mice were treated with buspirone (2 mg/kg, i.p.). The mice administered quercetin (5 mg/kg) spent a significantly longer percentage of time in the open arms of the EPM and their percentage of entries into the open arms was significantly increased compared to the vehicle-treated controls (p<0.05). The anxiolytic-like activities of quercetin were antagonized by trans-4-aminocrotonic acid (a $GABA_{A-{\rho}}$ agonist, 20 mg/kg) but not by flumazenil (a $GABA_A$ antagonist, 10 mg/kg) or WAY-100635 (a $5-HT_{1A}$ antagonist, 0.3 mg/kg). Moreover, there were no changes in the locomotor activity or myorelaxant effects in any group compared with the vehicle-treated controls. In the hole-board apparatus test, the number of head-dips increased significantly in the single treatment with quercetin (5 mg/kg) group compared to the vehicle-treated controls (p<0.05). These findings suggest that quercetin can promote anxiolytic-like activity, mediated by the GABAergic nervous system, in mice.