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Crystallization and X-Ray Crystallographic Studies of Wild-Type and Mutant Tryptophan Synthase α-Subunits from Escherichia coli

  • Jeong, Mi Suk;Jang, Se Bok
    • Molecules and Cells
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    • 제19권2호
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    • pp.219-222
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    • 2005
  • The a-subunit of Escherichia coli tryptophan synthase (${\alpha}TS$), a component of the tryptophan synthase ${\alpha}_2{\beta}_2$ complex, is a monomeric 268-residues protein (Mr = 28,600). ${\alpha}TS$ by itself catalyzes the cleavage of indole-3-glycerol phosphate to glyceraldehyde-3-phosphate and indole, which is converted to tryptophan in tryptophan biosynthesis. Wild-type and P28L/Y173F double mutant ${\alpha}$-subunits were overexpressed in E. coli and crystallized at 298 K by the hanging-drop vapor-diffusion method. X-ray diffraction data were collected to $2.5{\AA}$ resolution from the wild-type crystals and to $1.8{\AA}$ from the crystals of the double mutant, since the latter produced better quality diffraction data. The wild-type crystals belonged to the monoclinic space group C2 ($a=155.64{\AA}$, $b=44.54{\AA}$, $c=71.53{\AA}$ and ${\beta}=96.39^{\circ}$) and the P28L/Y173F crystals to the monoclinic space group $P2_1$ ($a=71.09{\AA}$, b=52.70, $c=71.52{\AA}$ and ${\beta}=91.49^{\circ}$). The asymmetric unit of both structures contained two molecules of ${\alpha}TS$. Crystal volume per protein mass ($V_m$) and solvent content were $2.15{\AA}^3\;Da^{-1}$ and 42.95% for the wild-type and $2.34{\AA}^3\;Da^{-1}$ and 47.52% for the double mutant.

생장점배양에 의한 우량마늘 체계적 증식 II 기내 인경 비대에 미치는 질소 및 Sucrose의 영향 (Systematic Propagation of High Quality Garlic (Allium sativum L.) Through Shoot Apical Meristem Culture II. Effects of Sucrose Concentration and Nitrogen Source on In Vitro formation of Bulblets)

  • Lee, Eun-Mo;Lee, Young-Bok
    • 식물조직배양학회지
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    • 제21권4호
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    • pp.193-200
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    • 1994
  • 마늘 생장점 배양을 통한 효율적인 우량종구 증식방법을 확립하기 위하여 기내배양시 유식물체 분화 및 기내인경 비대에 미치는 영향을 조사하였다. 유식물체 분화는 sucrose 및 질소 공급형태에 따라서 차이가 없었으나, 유식물체의 생장은 NO$_3$-N 공급시 저조하고, NH$_4$-N 및 NH$_4$+ NO$_3$ 공급형태에서 양호하였으며 특히 2차 계대배양시 질소형태를 바꾸워 공급할 경우 더욱 촉진되었다. 기내인경 비대는 NH$_4$-N를 공급하였을 때와 고농도의 Sucrose을 첨가하였을 때 촉진되었으며, 특히 유식물체에 저온처리를 하면 그 효과가 더 상승되었다. 기내배양에서 마늘의 생장에 이용되는 질소형태는 NH$_4$-N 이며, NO$_3$-N을 공급하면 흡수가 거의 되지 않고, $K^{+}$흡수도 저조하였다. Ethylene의 발생은 질소 공급 형태와 관계없이 8% sucrose 고농도에서 더 많이 발생되었다.

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만성 뇌졸중 환자들의 Sit to Stand Test의 임상적 유용성 (The Usability of Sit to Stand Test Performance in Chronic Stroke)

  • 조휘영;안승헌;이윤복;홍현화;이규창
    • 대한물리의학회지
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    • 제8권4호
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    • pp.549-558
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    • 2013
  • PURPOSE: This study is designed as a retrospective study, and identified the clinical usability of Sit to Stand (STS) test for predicting of fall incidence in stroke patients who experienced a fall within 1 year. METHODS: Between July 2011 and November 2012, 69 inpatients with stroke in K rehabilitation hospital were participated under voluntarily signing the informed consent form. STS test and 10m walk test (10MWT) were used to assess the muscle strength of lower-extremity and walking velocity, respectively. Also, we tested dynamic balance and motor function of lower-extremity in affected-side using with the Berg balance scale (BBS) and the Fugl-Meyer assessment of lower extremity (FM-L/E). METHODS: There were significant differences between subjects with fall-experienced group and without subjects without fall-experienced group in STS test, 10MWT, BBS scores and FM-L/E. STS test significantly showed a negative correlation between 10MWT (r=-.657), BBS (r=-.512), and FM-L/E (r=-.563). And, 10MWT have a influence on the performance of STS test (the capacity of explanation = 20%). The cut-off value of STS performance predicting falls experience is ${\geq}14.36$ seconds (sensitivity=76%; specificity=79%, area under curve=.785). According to logistic regression analysis of falls experience, subjects ${\geq}14.36$ s showed that 4.164 times (odd ratio) increased in falls than subjects < 14.36 s in STS test. CONCLUSION: This study demonstrated that STS test may be a useful tool predicting and measuring falls in patients with stroke. Further study will be needed to elucidate the kinematic analysis of STS test and the relationship between physical activity level and falls in stroke patients.

스테인리스강 STS 316L과 탄소강 A516-70의 이종금속 GTA 용접부 특성에 대한 연구 (A Study of Characteristics on the Dissimilar Metals (STS 316L - Carbon Steel: ASTM A516-70) Welds made with GTAW)

  • 김세철;신태우;문인준;장복수;고진현
    • Journal of Welding and Joining
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    • 제33권4호
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    • pp.37-43
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    • 2015
  • Characteristics of dissimilar metal welds between STS 316L and carbon steel ASTM A516 Gr.70 made with GTAW have been evaluated in terms of microstructure, ferrite content, chemical analysis, hardness and corrosion resistance. Three heat inputs of 9.00, 11.25, 13.00kJ/cm were employed to make joints of dissimilar metals with ER309 wire. Based on microstructural examination, the amount of vermicular type of ${\delta}$-ferrite was increased with increasing heat input due to the increase of Creq/Nieq in the second layer of welds. Based on the EDX analysis of weld metals, Cr and Ni content in the 2nd layer increased while those content in the first layer of welds decreased with heat inputs. Cellular solidification mode in the 1st layer and dendritic solidification mode in the 2nd layer due to different cooling rates were prevailed, respectively. Heat affected zone which formed hard microstructure showed higher hardness than the weld metal. The salt spray test of dissimilar metals weld joints showed that the carbon steel surfaces only corroded. The weight loss rate due to corrosion increased up to 100hours but it decreased above 100 hours. There was little difference in the weight loss caused by corrosion regardless of heat inputs.

정상성인 한국인에서 아스트로마이신 1회 점적 주사후 약물동태학적 평가 (Pharmacokinetics of Astromicin Following a Single Intravenous Infusion in Healthy Korean Subjects)

  • 복혜숙;최경업;김연화;백경란;송재훈
    • 한국임상약학회지
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    • 제13권2호
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    • pp.55-58
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    • 2003
  • Astromicin is an aminoglycoside antiviotic that is structually different from conventional aminoglycosides. Astromicin has been shown to be active against aerobic Gram-negative bacilli. The pharmacokinetics of astromicin were determined in 12 healthy volunteers ($65.5\pm5.23\;kg$ of body weight) following a 30-min continuous intravenous infusion at a dose of 200 mg. The plasma and urine samples were collected up to 24 h and drug concentrations were measured by a bioassay using Bacillus subtilis. Pharmacokinetic parameters were calculated by fitting individual concentration-time curve to a one-exponential decay model. The plasma levels were $16.9\pm1.68\;and\;1.05\pm0.346\l{\mu}g/ml$ at 0 h and 8 h after the infusion, respectively. The elimination half-life of astromicin was $1.86\pm0.360\;h$ The volume of distribution was $0.182\pm0.0164\;L/kg$, and the total body clearance was $5.25\pm1.74\;L/h$. These pharmacokinetic parameters were similar to these of gentamicin, tobramycin, and amikacin. Therefore, it is recommended that therapeutic drug monitoring of astromicin could be conducted in a similar fashion as the other aminoglycosides.

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낙동강 수계 주요 농업지대 소유역의 수질 오염 (Water Pollution in Some Agricultural Areas along Nakdong River)

  • 정종배;김복진;김정국
    • 한국환경농학회지
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    • 제16권2호
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    • pp.187-192
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    • 1997
  • 낙동강 수계의 주요 농업지대 소유역에 대한 수질조사 결과를 보면, 용존산소량, 전기전도도, 화학적산소요구량, pH, 중금속함량 등은 대부분 농업용수 기준에 비추어 볼 때 문제되지 않으나, 질소 함량은 전체적으로 농업용수 기준치를 상회하며 이온형태의 인은 특히 여름철에 대부분 지역에서 조류생장 최저 농도인 0.05mg/L 이상으로 측정되었다. 지하수의 경우 전기전도도는 하천수보다 높은 경향이고 질소 함량도 농업용수 기준을 훨씬 상회하였으며 지하수 중의 질산태 질소 함량은 음용수 수질 기준인 10mg/L를 초과하는 곳도 있었다. 지하수 중의 인 함량은 하천수와 비슷한 수준이었다. 중금속은 하천수에서와 마찬가지로 아연을 제외하고 납, 구리, 카드뮴 등은 거의 검출되지 않았다. 이상의 결과를 보면 조사 지역이 농업지대이므로 중금속 등의 요염은 문제되지 않지만 질소와 인에 의한 오염은 하천수와 지하수에서 상당한 수준에 달해 있으며 이러한 질소와 인의 오염은 결국 농경지에서의 화학비료와 퇴비의 시용, 축산폐수나 생활하수의 유입에 의한 것으로 보인다.

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Purification and Characterization of a 25 kDa Cathepsin L-like Protease from the Hemocyte of Coleopteran Insect, Tenebrio molitor Larvae

  • Jang, Kyung-Suk;Cho, Mi-Young;Choi, Hye-Won;Lee, Kang-Moon;Kim, Mi-Hee;Lee, Young-Un;Kurata, Shoichiro;Natori, Shunji;Lee, Bok-Luel
    • BMB Reports
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    • 제31권4호
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    • pp.364-369
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    • 1998
  • Insect plasma protein is abundant in the hemolymph of holometabolous insect larvae and is used as a source of amino acids and energy for construction of adult structures during metamorphosis. In order to understand the mechanism of decomposition of larval plasma proteins by hemocyte protease, we tried to purify a cysteine protease from the hemocyte lysate by using Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-Coumaryl-7-Amide (Z-Phe-Arg-MCA) as substrate and to identify plasma proteins that are selectively susceptible to the purified protease. Here, we describe the purification and characterization of a cysteine protease that specifically hydrolyzes the plasma protein of the coleopteran insect, Tenebrio molitor, larvae. The molecular mass of this enzyme was 25 kDa, as determined by SDS-PAGE under reducing conditions. The amino acids sequence of its $NH_{2}-terminus$ was determined to be Leu-Pro-Gly-Gln-Ile-Asp-Trp-Arg-Asp-Lys-Gly. This sequence contained Pro, Asp, and Arg residues, conserved in many papain superfamily enzymes. The specific cysteine protease inhibitors, such as E-64 and leupetin, inhibited its hydrolytic activity. One plasma protein with a molecular mass of 48 kDa was selectively hydrolyzed within 3 h when the purified enzyme and plasma proteins were incubated in vitro. However, the 48 kDa protein was not hydrolyzed by the purified 25 kDa protease in the presence of E-64. Western blotting analysis at various developmental stages showed that the purified enzyme was detected at larvae, pupae, and adult stages, but not the embryo stage.

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분극 시간에 따른 압전 페인트 센서의 압전 특성 연구 (Study on Piezoelectric Characteristics of Piezoelectric Paint Sensor According to Poling Time)

  • 한대현;박승복;강래형
    • 대한기계학회논문집A
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    • 제38권10호
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    • pp.1069-1074
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    • 2014
  • 본 논문에서는 분극 시간에 따라 압전 페인트 센서의 특성이 어떻게 변하는지 평가하고, 그 결과를 기술하였다. 연구에 사용된 압전 페인트 센서는 유연 압전 재료인 $Pb(Ni_{1/3}Nb_{2/3})O_3-Pb(Zr,Ti)O_2$ (PNN-PZT)와 에폭시 수지를 무게비 1:1로 혼합하여 제작하였다. 센서 시편은 몰드를 사용하여 $40{\times}10{\times}1mm^3$ 크기로 제작하였고, 윗면과 아랫면에 실버 페이스트를 사용하여 전극을 제작하였다. 분극 작업 시 온도는 상온으로, 분극 전계는 4kV/mm 고정한 상태에서 분극 시간을 달리함으로써 분극 조건을 달리하였다. 분극 특성은 충격 망치를 이용하여 충격을 시편에 가했을 때 충격 망치에서 측정되는 힘과 압전 페인트 센서에서 출력되는 전압을 비교하여 살펴보았다. 그 결과, 상온에서 분극 전계가 4kV/mm인 경우, 30분 정도 분극 처리를 한 경우 가장 최적의 분극 조건임을 확인하였다.

Bioequivalence of Cyclosporine A 100 mg Soft Capsules (Cipol-N® vs. Sandimmun Neoral®) in Healthy Korean Volunteers

  • Huh, Yong-Ho;Park, Eun-A;Chung, Youn-Bok;Pyo, Hee-Soo;Kwon, Oh-Seung
    • Journal of Pharmaceutical Investigation
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    • 제38권5호
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    • pp.343-348
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    • 2008
  • The bioequivalence of two cyclosporine A (CyA) 100 mg soft capsules (Chong Kun Dang's $Cipol-N^{(R)}$ as the test drug; Korea Novartis' Sandimmun $Neoral^{(R)}$ as the reference drug) was assessed in healthy male Korean volunteers after oral administration of 200 mg CyA according to a randomized crossover design. The whole blood samples were analyzed for the determination of parent CyA in the blood by using a validated HPLC/diode array detector method. The mean $AUC_t$ values for reference and test products were $4095.3{\pm}1397.2$ and $3958.3{\pm}1138.2\;ng{\cdot}hr/mL$, respectively. The mean $C_{max}$ values were $1135.9{\pm}293.2\;ng/mL$ for the reference product, and $985.0{\pm}207.9\;ng/mL$ for the test product. $T_{max}$ was $1.6{\pm}0.4\;hr$ for the reference and $1.8{\pm}0.5\;hr$ for the test product. The differences of $AUC_t$, $C_{max}$ and $T_{max}$ were -3.35, -13.28 and +10.63%, respectively. The point estimates and 90% confidence intervals for $AUC_t$ and $C_{max}$ were 0.981 (0.9171 to 1.0514) and 0.876 (0.8229 to 0.9336), respectively. Based on the pharmacokinetic and statistical data, we conclude that these two products are bioequivalent and can be considered interchangeable in the medical practice.

레티노산 함유 폴리락탄산 나노입자의 제조 및 약물 방출 (Preparation and Drug Release of All-Trans Retinoic Acid-Loaded Poly(L-lactic acid) Nanoparticles)

  • 채지만;이경만;김인숙;이용복;신상철;오인준
    • Journal of Pharmaceutical Investigation
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    • 제34권5호
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    • pp.401-406
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    • 2004
  • To develop an intravenous delivery system of all-trans retinoic acid (ATRA) for the cancer therapy, poly(L-lactic acid) nanoparticles were prepared and characterized. Emulsification-solvent evaporation method was chosen to prepare submicron sized nanoparticles. Spherical nanoparticles less than 200 nm in diameter with narrow size distribution were prepared, and the entrapment efficiency of drug was more than 95%. The endothermic peak at $183^{\circ}C$ and X-ray crystallographic peak of ATRA appeared in the nanoparticle system, suggesting the inhibition of crystallization of ATRA by polymer adsorption during the precipitation process. ATRA was released at $37^{\circ}C$ for 60 days and the release rate was dependent on the concentration of drug incorporated in the nanoparticles. While ATRA was unstable in the light, it was very stable at $4^{\circ}C$. These results suggest the usefulness of PLA nanoparticles as a sustained and prolonged release carrier for ATRA.