• 제목/요약/키워드: Rapid kinetics

검색결과 133건 처리시간 0.028초

Cholesterol Biosynthesis from Lanosterol: Development of a Novel Assay Method, Characterization, and Solubilization of Rat Hepatic Microsomal Sterol Δ7-Reductase

  • Lee, Joon-No;Paik, Young-Ki
    • BMB Reports
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    • 제30권5호
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    • pp.370-377
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    • 1997
  • A novel assay method is described for rapid quantitation of reaction rate of sterol ${\Delta}^7$-reductase (${\Delta}^7$-SR) which catalyzes reduction of the ${\Delta}^7$-double bond of sterols. Of six different organ tissues-liver, small intestine, brain, lung, kidney, and testis-. ${\Delta}^7$-SR activity was detected only in liver (2.30 nmol/min/mg protein) and testis (0.11 nmol/min/mg protein). Using a newly developed method which employs diet-induced enzyme proteins and ergosterol as substrate, we assessed both kinetics ($K_m=210\;{\mu}M$, $V_{max}=1.93\;nmol/min/mg$) and inhibition of the rat hepatic ${\Delta}^7$-SR against well-studied cholesterol lowering agents such as triparanol ($IC_{50}=16\;{\mu}M$). 3-$\beta$-[2-(diethylamino)ethoxy]androst-5-en-17-one (U18666A) ($IC_{50}=5.2\;{\mu}M$), and trans-1.4-bis(2-chlorobenzylaminomethyl)cyclohexane dihydrochloride (AY-9944) ($IC_{50}=0.25\;{\mu}M$). Of the three well-known AY-9944-sensitive cholesterogenic enzymes (i.e., ${\Delta}^7$-SR, sterol ${\Delta}^8$-isomerase, and sterol ${\Delta}^14$-reductase). ${\Delta}^7$-SR was found to be the most sensitive enzyme with a noncompetitive inhibition of this compound ($K_i=0.109\;{\mu}M$). Substrate specificity studies of the microsomal ${\Delta}^7$-SR indicate that the relative reaction rate for 7-dehydrocholesterol and ergosterol are 5.6-fold and 1.6-fold higher than that for lathosterol. ${\Delta}^7$-SR activity was also modulated by feeding rats a diet supplemented with 0.5% ergosterol (>2.6-fold) in addition to 5.0% cholestyramine plus 0.1% lovastatin ($\simeq$5.0-fold). Finally, microsomal ${\Delta}^7$-SR was solubilized by 1.5% 3-[3-(cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) and enriched on PEG (0~10%) precipitation, which should be suitable for further purification of the enzyme.

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Photocatalytic Oxidation of 2-Mercaptoethanol to Disulfide using Sb(V)-, P(V)-, and Ge(IV)-porphyrin Complexes

  • Shiragami, Tsutomu;Onitsuka, Dai;Matsumoto, Jin;Yasuda, Masahide
    • Rapid Communication in Photoscience
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    • 제3권4호
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    • pp.70-72
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    • 2014
  • Visible-light irradiation of MeCN solution containing di(hydroxo)metallo(tetraphenyl)porphyrin complex $(tppM(OH)_2$: 1a; $M=Sb(V)^+Br^-$, 1b; $M=P(V)^+Cl^-$, 1c; M=Ge(IV)) and 2-mercaptoethanol (2-ME) as a substrate under aerated condition gave bis(2-hydroxyethyl)disulfide (2-HEDS) as an oxidative product of 2-ME. It is indicated that the oxidation of 2-ME should proceed with a photocatalytic process by 1, because the turn over number (TON) for the formation of 2-HEDS was over unit. The TON was determined to be 642 as a maximum value when 1a was used as a sensitizer. The formation of 2-HDES was extremely slow under argon atmosphere. The fluorescence of 1 was not quenched by 2-ME at all, and the free energy change (${\Delta}G$) with electron transfer (ET) from 2-ME to excited triplet state of $1(^31^*)$ was estimated as a negative value. The quenching rate constant ($k_r$) of $^31^*$ by 2-ME, obtained by the kinetics for the formation of 2-HEDS, strongly depends on ${\Delta}G$. These findings indicate that 1-sensitized oxidation was initiated by photoinduced ET from 2-ME to $^31^*$ to generate both radical cation of 2-ME ($2-ME^{+\bulle}$) and porphyrin radical anion ($1^{-\bulle}$), resulting that the formation of 2-HEDS can be proceeded by the dimerization of $2-ME^{+\bulle}$, and through a catalytic cycle due to returning to 1 by the ET from $1^{-\bulle}$ to molecular oxygen.

Buccal Delivery of [D-Ala2, D-Leu5]Enkephalin Incorporated in Mucoadhesive Poly(acrylic acid) Hydrogels

  • Lee, Jae-Hwi;Lee, Yoon-Jin;Kang, Kyoung-Hoon;Nam, Dae-Young;Choi, Young-Wook
    • Journal of Pharmaceutical Investigation
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    • 제35권5호
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    • pp.369-373
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    • 2005
  • The objectives of the current work is to understand the factors impacting the formulation and performance of a Carbopol mucoadhesive buccal delvery system for a model peptide drug, $[D-Ala{^2},\;D-Leu{^5}]$enkephalin (DADLE, Mw=569.7) with comparable chemical and enzymatic stability. Specifically, in vitro buccal DADLE delivery from the cross-linked poly(acrylic acid) (PAA) hydrogel system was characterized. In addition, the influences of several penetration enhancers on the ex vivo buccal absorption of DADLE were also studied. In this study, the PAA hydrogels generally swell to 100% of their original weight in the phosphate pH 7.4 buffer. The water penetration into the PAA hydrogel occurred based on a zero-order kinetics for the first 60 min and steadily decreased afterwards. From the release study, it can be seen that the initial DADLE release was so rapid and the rate of release of DADLE decreased as the time elapsed. The porcine buccal tissue was found to be permeable to DADLE with a flux value of $0.07%/cm{^2}/hr({\pm}0.01\;SD)$. From the ex vivo diffusion study, it was found that sodium taurodihydrofusidate showed a greater degree of enhancement compared to the phospholipids with an Enhancement Ratio (ER) of 8.7 compared to 2.7 and 1.9 for didecanoylphosphatidylcholine and lysophosphatidylcholine, respectively. The work encompassed within this paper has demonstrated the feasibility of using the PAA hydrogel delivery system with its good mucoadhesive properties for the buccal delivery of peptides.

Inhibitory and Excitatory Postsynaptic Currents of Medial Vestibular Nucleus Neurons of Rats

  • Chun, Sang-Woo;Choi, Jeong-Hee;Park, Byung-Rim
    • The Korean Journal of Physiology and Pharmacology
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    • 제7권2호
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    • pp.59-63
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    • 2003
  • The medial vestibular nucleus (MVN) neurons are controlled by excitatory synaptic transmission from the vestibular afferent and commissural projections, and by inhibitory transmission from interneurons. Spontaneous synaptic currents of MVN neurons were studied using whole cell patch clamp recording in slices prepared from 13- to 17-day-old rats. The spontaneous inhibitory postsynaptic currents (sIPSCs) were significantly reduced by the $GABA_A$ antagonist bicuculline ($20{\mu}M$), but were not affected by the glycine antagonist strychnine ($1{\mu}M$). The frequency, amplitude, and decay time constant of sIPSCs were $4.3{\pm}0.9$ Hz, $18.1{\pm}2.0$ pA, and $8.9{\pm}0.4$ ms, respectively. Spontaneous excitatory postsynaptic currents (sEPSCs) were mediated by non-NMDA and NMDA receptors. The specific AMPA receptor antagonist GYKI-52466 ($50{\mu}M$) completely blocked the non-NMDA mediated sEPSCs, indicating that they are mediated by an AMPA-preferring receptor. The AMPA mediated sEPSCs were characterized by low frequency ($1.5{\pm}0.4$ Hz), small amplitude ($13.9{\pm}1.9$ pA), and rapid decay kinetics ($2.8{\pm}0.2$ ms). The majority (15/21) displayed linear I-V relationships, suggesting the presence of GluR2-containing AMPA receptors. Only 35% of recorded MVN neurons showed NMDA mediated currents, which were characterized by small amplitude and low frequency. These results suggest that the MVN neurons receive excitatory inputs mediated by AMPA, but not kainate, and NMDA receptors, and inhibitory transmission mediated by $GABA_A$ receptors in neonatal rats.

Robust power control design for a small pressurized water reactor using an H infinity mixed sensitivity method

  • Yan, Xu;Wang, Pengfei;Qing, Junyan;Wu, Shifa;Zhao, Fuyu
    • Nuclear Engineering and Technology
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    • 제52권7호
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    • pp.1443-1451
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    • 2020
  • The objective of this study is to design a robust power control system for a small pressurized water reactor (PWR) to achieve stable power operations under conditions of external disturbances and internal model uncertainties. For this purpose, the multiple-input multiple-output transfer function models of the reactor core at five power levels are derived from point reactor kinetics equations and the Mann's thermodynamic model. Using the transfer function models, five local reactor power controllers are designed using an H infinity (H) mixed sensitivity method to minimize the core power disturbance under various uncertainties at the five power levels, respectively. Then a multimodel approach with triangular membership functions is employed to integrate the five local controllers into a multimodel robust control system that is applicable for the entire power range. The performance of the robust power system is assessed against 10% of full power (FP) step load increase transients with coolant inlet temperature disturbances at different power levels and large-scope, rapid ramp load change transient. The simulation results show that the robust control system could maintain satisfactory control performance and good robustness of the reactor under external disturbances and internal model uncertainties, demonstrating the effective of the robust power control design.

토양 훈증제 1,3-Dichloropene의 물 및 토양 중 분해 (Transformation for 1,3-Dichloropene of Soil Fumigant in Water and Soil)

  • 김정호
    • 한국환경과학회지
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    • 제16권12호
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    • pp.1463-1468
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    • 2007
  • Emission of methyl bromide(MeBr) of soil fumigant was implicated in stratospheric ozone depletion. To determine the environmental fate for 1,3-dichloropene(1,3-D) of alternatives fumigants for MeBr, this paper researched the transformation for 1,3-D in water and soil. Half lives of cis-1,3-D in water with first-order kinetics are 9.9day and 1.7day at $25^{\circ}C\;and\;40^{\circ}C$, half lives of trans-1,3-D are 8.6day and 1.5day at $25^{\circ}C\;and\;40^{\circ}C$, respectively. Transformation for 1,3-D in water at high temperature faster then at low temperature. Hydrolysis for 1,3-D in water are unaffected at $pH\;2.5{\sim}pH\;10.0$, but hydrolysis for 1,3-D at pH 11.5 higher then at $pH\;2.5{\sim}pH\;10.0$. Half lives of cis-1,3-D in soil are 11.5day and 7.7day at 3% and 10% of soil moisture, half lives of trans-1,3-D are 9.9day and 6.9day at 3% and 10% of soil moisture, respectively. Transformation for 1,3-D in water increased with increasing soil moisture. Transformation for trans-1,3-D isomer are more rapid then cis-1,3-D isomer in water and soil. This research has identified that transformation for 1,3-dichloropropene are affected by temperature, pH, soil moisture, and isomer of cis and trans in water and soil.

Affinity Labeling of E. coli GTP Cyclohydrolase I by a Dialdehyde Derivative of Guanosine Triphosphate

  • Ahn, Chi-Young;Park, Sang-Ick;Kim, Ju-Myeong;Yim, Jeong-Bin
    • BMB Reports
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    • 제28권1호
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    • pp.72-78
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    • 1995
  • Time-dependent inactivation of E. coli GTP cyclohydrolase I with a 2',3'-dialdehyde derivative of GTP (oGTP) was directed to the active site of the enzyme, and was dependent on the concentration of oGTP. The kinetics of inactivation were biphasic with a rapid reaction occurring immediately upon exposure of the enzyme to oGTP followed by a slow rate of inactivation. The $K_i$ value of oGTP for the enzyme was 0.25 mM. Inactivation was prevented by preincubation of the enzyme with GTP, the substrate of the enzyme. At 100% inactivation, 2.3 mol of [8.5'-$^3H$]oGTP were bound per each enzyme subunit, which consists of two identical polypeptides. The active site residue which reacted with the affinity label was lysine. oGTP interacted selectively with the ${\varepsilon}$-amino group of lysine in the GTP-binding site to form a morpholine-like structure which was stable without sodium borohydride treatment. However, triphosphate group was eliminated during the hydrolysis step. To identify the active site of the enzyme, [8.5'-$^3H$]oGTP-labeled enzyme was cleaved by endoproteinase Lys-C, and the $^3H$-labeled peptide was purified by HPLC. The amino acid sequence of the active site peptide was Pro-Ser-Leu-Ser-Lys, which corresponds to the aminoterminal sequence of the enzyme.

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합성가스(H2/CO) 예혼합 충돌 제트화염에서 신장률에 따른 부상된 화염 구조에 관한 연구 (A Study on the Lifted Flame Structure with Strain Rates in Premixed Impinging Jet Flames of Syngas (H2/CO))

  • 심근선;장병록;이기만
    • 한국수소및신에너지학회논문집
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    • 제26권4호
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    • pp.347-356
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    • 2015
  • A study has been conducted numerically to investigate the lifted flat syngas flame structure of impinging jet flame configuration with the global strain rates in 10% hydrogen content. In this study, the effects of strain rate were major parameters on chemistry kinetics and flame structure at stagnation point. The numerical results were calculated by SPIN application of the CHEMKIN package. The strain rates were adjusted with Reynolds numbers of premixed syngas-air mixture. Different flame shapes were observed with different strain rates. As strain rate has increased, the flame temperature and axial velocity have been decreased due to the flame heat loss increment, and the OH radical reaction zones become narrower but each mole fractions are still constant. Also, the reversion of $H_2O$ product near stagnation point has been found out when strain rate has increased. This phenomenon is attributed to the rapid production of oxidizing radical reaction such as the R12 ($H+O_2(+M)=HO_2(+M)$), which makes the R18 ($HO_2+OH=O_2+H_2O$) reaction increment.

박테리오파지 f2에 대한 자외광선의 살균효과 (The Inactivation Effects of UV Light on Bacteriophage f2)

  • Kim, Chi-Kyung;Quae Chae
    • 한국미생물·생명공학회지
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    • 제11권3호
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    • pp.155-161
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    • 1983
  • Bacteriophage f2에 대한 자외광선의 살균효과와 외투막 단백질의 구조에 미치는 영향을 Ray-onet photoreactor PPR-208을 사용하여 300nm의 광선으로 연구하였다. 처음 20분간의 조사에서는 약 4 log의 phage가 감소되고 그후 완만한 살균효과를 보이다가 90분 이상의 조사에서는 생존 바이러스가 발견되지 않았다. Tryptophan residue의 fluorescenve quenching, 자외선으로 조신한 phage에 부착시킨 ANS (8-anilino-1-napht-halene sulfonate)의 fluorescence emission의 감소, tryptophan에서 ANS로의 energy transfer 의 감소 등 spectroscopic technique에 의한 결과와 자외선 조사에 의하여 단백질 외투만이 파손되는 전자현미경 관찰의 결과에 의하여 자외광선은 phage f2의 외투막 단백질의 구조에 변화를 일으킨다는 것이 밝혀졌다.

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Fabrication and Characterization of Dye-sensitized Solar Cells based on Anodic Titanium Oxide Nanotube Arrays Sensitized with Heteroleptic Ruthenium Dyes

  • Shen, Chien-Hung;Chang, Yu-Cheng;Wu, Po-Ting;Diau, Eric Wei-Guang
    • Rapid Communication in Photoscience
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    • 제3권1호
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    • pp.16-19
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    • 2014
  • Anodic self-organized titania nanotube (TNT) arrays have a great potential as efficient electron-transport materials for dye-sensitized solar cells (DSSC). Herewith we report the photovoltaic and kinetic investigations for a series of heteroleptic ruthenium complexes (RD16-RD18) sensitized on TNT films for DSSC applications. We found that the RD16 device had an enhanced short-circuit current density ($J_{SC}/mAcm^{-2}=15.0$) and an efficiency of power conversion (${\eta}=7.2%$) greater than that of a N719 device (${\eta}=7.1%$) due to the increasing light-harvesting and the broadened spectral features with thiophene-based ligands. However, the device made of RD17 (adding one more hexyl chain) showed smaller $J_{SC}(14.1mAcm^{-2})$ and poorer ${\eta}(6.8%)$ compare to those of RD16 due to smaller amount of dye-loading and less efficient electron injection for the RD17 device than for the RD16 device. For the RD18 dye (adding one more thiophene unit and one more hexyl chain), we found that the device showed even lower $J_{SC}(13.2mAcm^{-2}) $ that led to a poorest device performance (${\eta}=6.2%$) for the RD18 device. These results are against to those obtained from the same dyes sensitized on $TiO_2$ nanoparticle films and they can be rationalized according to the electron transport kinetics measured using the methods of charge extraction and transient photovoltage decays.