• Title/Summary/Keyword: KD-$(k_0,\

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Pharmacological Characterization of (10bS)-1,2,3,5,6,10b-hexahydropyrrolo[2,1-a]isoquinoline Oxalate (YSL-3S) as a New ${\alpha}_2$-Adrenoceptor Antagonist

  • Chung, Sung-Hyun;Yook, Ju-Won;Min, Byung-Jun;Lee, Jae-Yeol;Lee, Yong-Sup;Jin, Chang-Bae
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.353-359
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    • 2000
  • ${\alpha}_2$-Adrenoceptor antagonists, which can enhance synaptic norepinephrine levels by blocking feedback inhibition processes, are potentially useful in the treatment of disease states such. as depression, memory impairment, impotence and sexual dysfunction. (10bS)-1,2,3,5,6,10b-Hexahydropyrrolo[2,1-a]isoquinoline oxalate (YSL-3S) was evaluated in several in vitro biological tests to establish its pharmacological profile of activities as an ${\alpha}_2$-adrenoceptor antagonist. Saturation binding assay revealed that$^{3}[H]$rauwolscine bound to the $\alpha$$_2$-adrenoceptors with a Kd value of 6.3$\pm$0.5 nM and a Bmax value of 25l$\pm$39 fmol/mg protein in rat cortical synaptic membranes. Competitive binding assay showed that YSL-3S inhibited the binding of$^3[H]$rauwolscine (1 nM) in a concentration-dependent manner with a Ki value of 98.2$\pm$12.1 nM while it did not inhibit the binding of [$^3$H]cytisine (1.25 nM) to neuronal nicotinic cholinergic receptors. The Ki values of yohimbine, clonidine and norepinephrine for $^3[H]$rauwolscine binding were 15.8$\pm$1.0, 40.1$\pm$5.9 and 40.0$\pm$11.5 nM, respectively. In addition, the binding affinity of YSL-3S for ${\alpha}_2$-adrenoceptors was higher than that of its antipode and the racemic mixture. The functional activity of YSL-3S at the presynaptic ${\alpha}_2$-adrenoceptors was assessed using the prostatic portion of the rat vas deferens. Clonidine inhibited field-stimulated contractions of the vas deference in a dose-dependent manner. The presence of YSL-3S or yohimbine caused a parallel, rightward the dose-response curve of clonidine in a dose-dependent manner, indicating an antagonistic action at the presynaptic ${\alpha}_2$-adrenoceptors. The $pA_2$values of yohimbine and YSL-3S were 7.66$\pm$0.13 and 6.64$\pm$0.18, respectively. The results indicate that YSL-3S acts as a competitive antagonist at presynaptic ${\alpha}_2$ -adrenoceptors with a potency approximately ten times lower than yohimbine, but is devoid of binding affinity for neuronal nicotinic cholinergic receptors.

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Influence of Ph and Temperature on Polyphenol Oxidase in the Leaves of Perilla frutescens var. japonica (들깨잎 폴리페놀 산화효소의 pH 및 온도에 의한 영향)

  • Kim, Yoo-Kyung;Kim, An-Keun
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.384-390
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    • 2004
  • Polyphenol oxidase-catalyzed oxidation of substrates (t-butylcatechol, 4-methylcatechol, chlorogenic acid, caffeic acid and pyrocatechol) were performed in the Ph range 4~8. Co ncentrations of substrate's major oxidation products were monitored by high performance liquid chromatograph. The nature and amounts of products formed were highly pH dependent. They also were ifluenced by kinds of substrates. Major oxidation product of 4-methylcatechol appeared the maxium value at pH 5, them of chlorogenic acid, caffeic acid and pyrocatechol at pH 6.0 and that of t-butylcatechol at pH 5~7. Time-dependent PPO activity was determined at $4^{\circ}C\;and\;30^{\circ}C$. PPO extracted by phosphate buffer containing triton X-114 (t-PPO) was more stable than PPO by phosphate buffer (b-PPO). The result of electrophoresis, at first PPO was showed only a band at 48 kd. After 1~3 days a partial degrade band was appeared in b-PPO and three partial degrade bands in t-PPO. No activity band was appeared in PPOs at $30^{\circ}C$ and b-PPO at $4^{\circ}C$ after 4 days. And a band (37 kDa) in t-PPO was remained finally and disappered. PPO from Perillae leaves has two activity bands at 48 and 37 kDa in previous paper. It was supposed that PPO in the leaves of Perilla frutescens was a protein having one molecular weight as 48 kDa. And 37 kDa protein, relatively proteolysis-resistant, was a proteolyzed form of a major form.

Through Knee Amputation: Technique Modifications and Surgical Outcomes

  • Albino, Frank P.;Seidel, Rachel;Brown, Benjamin J.;Crone, Charles G.;Attinger, Christopher E.
    • Archives of Plastic Surgery
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    • v.41 no.5
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    • pp.562-570
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    • 2014
  • Background Knee disarticulations (KD) are most commonly employed following trauma or tumor resection but represent less than 2% of all lower extremity amputations performed in the United States annually. KDs provide enhanced proprioception, a long lever arm, preservation of adductor muscle insertion, decreased metabolic cost of ambulation, and an end weight-bearing stump. The role for KDs in the setting of arterial insufficiency or overwhelming infection is less clear. The purpose of this study is to describe technique modifications and report surgical outcomes following KDs at a high-volume Limb Salvage Center. Methods A retrospective study of medical records for all patients who underwent a through-knee amputation performed by the senior author (C.E.A.) between 2004 and 2012 was completed. Medical records were reviewed to collect demographic, operative, and postoperative information for each of the patients identified. Results Between 2004 and 2012, 46 through-knee amputations for 41 patients were performed. The mean patient age was 68 and indications for surgery included infection (56%), arterial thrombosis (35%), and trauma (9%). Postoperative complications included superficial cellulitis (13%), soft tissue infection (4%), and flap ischemia (4%) necessitating one case of surgical debridement (4%) and four transfemoral amputations (9%). 9 (22%) patients went on to ambulate. Postoperative ambulation was greatest in the traumatic cohort and for patients less than 50 years of age, P<0.05. Alternatively, diabetes mellitus and infection reduced the likelihood of postoperative ambulation, P<0.01. Conclusions Knee disarticulations are a safe and effective alternative to other lower extremity amputations when clinically feasible. For patient unlikely to ambulate, a through-knee amputation maximizes ease of transfers, promotes mobility by providing a counterbalance, and eliminates the potential for knee flexion contracture with subsequent skin breakdown.

Strategic construction of mRNA vaccine derived from conserved and experimentally validated epitopes of avian influenza type A virus: a reverse vaccinology approach

  • Leana Rich Herrera-Ong
    • Clinical and Experimental Vaccine Research
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    • v.12 no.2
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    • pp.156-171
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    • 2023
  • Purpose: The development of vaccines that confer protection against multiple avian influenza A (AIA) virus strains is necessary to prevent the emergence of highly infectious strains that may result in more severe outbreaks. Thus, this study applied reverse vaccinology approach in strategically constructing messenger RNA (mRNA) vaccine construct against avian influenza A (mVAIA) to induce cross-protection while targeting diverse AIA virulence factors. Materials and Methods: Immunoinformatics tools and databases were utilized to identify conserved experimentally validated AIA epitopes. CD8+ epitopes were docked with dominant chicken major histocompatibility complexes (MHCs) to evaluate complex formation. Conserved epitopes were adjoined in the optimized mVAIA sequence for efficient expression in Gallus gallus. Signal sequence for targeted secretory expression was included. Physicochemical properties, antigenicity, toxicity, and potential cross-reactivity were assessed. The tertiary structure of its protein sequence was modeled and validated in silico to investigate the accessibility of adjoined B-cell epitope. Potential immune responses were also simulated in C-ImmSim. Results: Eighteen experimentally validated epitopes were found conserved (Shannon index <2.0) in the study. These include one B-cell (SLLTEVETPIRNEWGCR) and 17 CD8+ epitopes, adjoined in a single mRNA construct. The CD8+ epitopes docked favorably with MHC peptidebinding groove, which were further supported by the acceptable ∆Gbind (-28.45 to -40.59 kJ/mol) and Kd (<1.00) values. The incorporated Sec/SPI (secretory/signal peptidase I) cleavage site was also recognized with a high probability (0.964814). Adjoined B-cell epitope was found within the disordered and accessible regions of the vaccine. Immune simulation results projected cytokine production, lymphocyte activation, and memory cell generation after the 1st dose of mVAIA. Conclusion: Results suggest that mVAIA possesses stability, safety, and immunogenicity. In vitro and in vivo confirmation in subsequent studies are anticipated.

Biochemical Characterization of an Extracellular Protease in Serratia proteamaculans Isolated from a Spider (무당거미에서 분리한 Serratia proteamaculans에서 분비되는 단백질분해효소의 생화학적 특성)

  • Lee Kieun;Kim Chul-Hee;Kwon Hyun-Jung;Kwak Jangyul;Shin Dong-Ha;Park Doo-Sang;Bae Kyung-Sook;Park Ho-Yong
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.269-274
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    • 2004
  • Serratia proteamaculans isolated from the midgut of a spider formed big halos around the bacterial colonies, indicating that the bacterial strain produces an extracellular protease. Activity staining of the extracellular pro­tein fractions using zymogram also demonstrated that the major protein with an estimated molecular mass of 52 kDa contained a high proteolytic activity. The protease was purified to near electrophoretic homogeneity from the culture supernatant after filtration and ion exchange and size exclusion chromatography. The purified enzyme had a relatively high proteolytic activity between pH 6.0 and 10.0 and at broad temperature range. The proteolytic activity of the enzyme was not inhibited by phenylmethylsulfonyl fluoride but strongly inhibited by 1, 10-phenanthroline and EDTA. The activity also was dependent on the presence of $Ca^{++}\;and\;Zn^{++}$ ions. These observations indicate that the enzyme is a metalloprotease.

Purification of Mitochondrial Matrix Aldehyde Dehydrogenase from Pig Brain

  • Kim, Kyu-Tae;Lee, Young-Don
    • BMB Reports
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    • v.28 no.2
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    • pp.177-183
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    • 1995
  • The activity of aldehyde dehydrogenase (ALDH) in the cerebrum, cerebellum, striatum, and medulla oblongata was examined and mitochondrial matrix ALDH was purified prior to immunohistochemical study on the localization of ALDH isozymes in pig brain. Relatively high enzyme activity was found in the striatum and medulla oblongata when using indole-3-acetaldehyde as substrate, and in the striatum when using 3,4-dihydroxyphenylacetaldehyde (DOPAL). The main part of mitochondrial ALDH activities with both acetaldehyde and DOPAL existed in the matrix fraction. The ratio of activity of the matrix to the membrane fraction in the cerebrum was higher than in the cerebellum, suggesting that the distribution pattern of ALDH isozymes was different according to the brain regions. The 276-fold purified mitochondrial matrix ALDH from pig brain was identified to be homologous tetramers with 53 KD subunits. The enzyme showed maximal activity at pH 9.0 and was stable in the temperature range from $25^{\circ}C$ to $37^{\circ}C$. The mitochondrial matrix ALDH activity was considerably inhibited by acetaldehyde in vitro. The $K_m$ values of the enzyme for acetaldehyde and propionaldehyde were 5.8 mM and 4.9 mM, respectively, whereas $K_m$ values for indole-3-acetaldehyde and DOPAL were 44 ${\mu}M$ and 1.6 ${\mu}M$, respectively. The $V_{max}/K_{m}$ ratio was the highest with DOPAL as compared with other substrates. These results suggested that mitochondrial matrix ALDH in the present work might be a low Km isozyme involved in biogenic aldehyde oxidation in pig brain.

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Impulse Breakdown Behaviors of Dry Air as an Alternative Insulation Gas for SF6

  • Li, Feng;Yoo, Yang-Woo;Kim, Dong-Kyu;Lee, Bok-Hee
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.25 no.3
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    • pp.24-32
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    • 2011
  • [ $SF_6$ ]gas, which has an excellent dielectric strength and interruption performance, is used in various applications such as gas insulated switchgear (GIS) in substations. However, since $SF_6$ has a high global warming potential (GWP), it is necessary to find an eco-friendly alternative insulation gas. In order to examine the possibility of using alternative insulation gases for $SF_6$ in power distribution system equipment, the dielectric strength and physical phenomena of dry air in a quasi-uniform electric field are investigated experimentally in this paper. As a result, the breakdown voltages for positive polarity are higher than those for negative polarity under impulse voltage applications. The negative 50[%] flashover voltage, $V_{50}$ of dry air under conditions above 0.4[MPa] gas pressure, is higher than 150[kV], that is the basic impulse insulation level of distribution equipment. The $V_{50}$ increases linearly with increasing the gas pressure, regardless of the waveform and polarity of the applied impulse voltages. The voltage-time curves are dependent on the rise time of the impulse voltage and gas pressure. Furthermore, streamer discharge was observed through light emission images by an ICCD camera under impulse voltage applications.

Preliminary study on colloidal partitioning and speciation of trace metals in acid mine drainage

  • Kwon, Jang-Soon;Lee, Jeong-Ho;Yun, Seong-Taek;Jung, Hun-Bok;Chang, Min-Kyoung;Lee, Pyeong-Ku
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.100-101
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    • 2004
  • Many researches in Korea have been performed to understand the pollution of stream waters by acid mine drainage. However, few studies have been conducted regarding the effect of particulate and colloidal fractions on the transport of trace metals. To estimate harmful effects of trace metals, it is important to evaluate the particulate and colloidal metals as well as dissolved metals, because particulate and colloidal fractions of trace metals play an important role in transport of trace metals and may adversely affect habitats and organisms in riverine system. Colloids are solids with effective diameters in size range from 0.001 $\mu$m to 1 $\mu$m. According to Jone et al. (1974), metals in surface water, like Al, Fe, and Mn, require filtration with pore-size membranes smaller than 0.45 $\mu$m to define dissolved concentrations. The main objective of this study is to understand the effects of particulate, colloidal, and truly dissolved fractions on the transport and fate of trace metals in acid mine drainage. This study was conducted for the Onjeong creek in the Uljin mine area. Sampling was carried out in 13 sites, spatially covering the area from mine dumps to the downstream Onjeong reservoir. To examine the metal partitioning between particulate, colloidal, and truly dissolved fraction, we used successive filtration techniques consisting of conventional method (using 0.45 $\mu$m membranes) and tangential-flow ultrafiltration (using 0.001 $\mu$mm membranes). Ultrafiltration may seperate much smaller particles from aqueous phase (Josephson, 1984; Hernandez and Stallard, 1988). The analysis of metals were performed by inductively coupled plasma - atomic emission spectrometer (ICP-AES: model Perkin Elmer OPTIMA3000XL). Anions such as SO$_4$, Cl and NO$_3$ were measured with ion chromatograph (IC: model Dionex 120). Sample analysis is still in progress. The preliminary data show that the studied creek is severely polluted by Al, Fe, Mn, Pb and Zn. Toward upstream sites with relatively lower pH, less than 50% of Al and Fe occur in the sorbed form on particles or colloids, whereas more than 80% of Al and Fe occur in the sorbed form in downstream sites or tributaries with relatively higher pH. Less than 30% of Zn is present in particle or colloidal forms in the whole range of creek. Truly dissolved fraction of trace metals is negatively correlated with pH. The Kd values for Al, Fe and Zn consistently increase with increasing pH and decrease with increasing particle concentration.

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Codon usage analysis of rice glutelin genes (쌀 저장 단백질 글루텔린 유전자 암호 분석)

  • Shin, Yun-Cheol;Kim, Ju-Kon;Nahm, Baek-Hie
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.517-524
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    • 1993
  • To characterize glutelins, the most abundant storage protein in rice, 13 complete coding sequences of glutelin genes from the database were analyzed. According to the phylogenic analysis, these genes could be classified into 5 groups, Group I to V. The degrees of homology were calculated to be in the range of 90 to 60%, but the patterns of hydrophobicity were similar in all the groups. Also, each group was found to have similar amino acid composition with variations in lysine content from 2.5 to 3.6% due to the point mutation of arginine to lysine. The isoelectric points of mature proteins and their basic chains of all the groups showed the value of about 9.0 and 10.0, respectively, while the isoelectric points of acidic chains in these groups showed the distinct value of 6.6, 6.7, 7.2, 8.4 and 7.9. The plot of the fraction of G+C at synonimous site in codons (GC3s) against effective codon numbers suggest no major difference in translational efficiency in the expression of glutelin multigenes.

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Ion Exchange Characteristics of Novel HDBPDA and Dowex Ion Exchange Resins (새로운 HDBPDA와 Dowex 이온교환수지의 이온교환 특성)

  • Kim, Dong Won;Kim, Chang Suk;Choi, Ki Young;Jeon, Young Shin;Jeong, Young Kyu;Yoon, Yeo Hag;Hon, Choon Pyo
    • Analytical Science and Technology
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    • v.6 no.5
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    • pp.463-469
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    • 1993
  • HDBPDA ion exchange resin, {(4, 5) : (13, 14)-Dibenzo-6, 9, 12-trioxa-3, 15, 21, -triazabicyclo [15. 3. 1]heneicosa-(1, 17, 19)(18, 20, 21) triene ion exchange resin : HDBPDA ion exchange resin} had a capacity of 3.8meq/g dry resin. The distribution coefficients of alkali and alkaline earth metal ions on HDBPDA and strongly acidic cation exchange resin, Dowex 50W-X8(200-400mesh) in water, and the various concentrations of hydrochloric acid were determined. Concentration of hydrochloric acid have almost not influenced on the distribution coefficients of alkali and alkaline earth metal ions for the HDBPDA ion exchange resin, but generally the distribution coefficient slightly increased with decreasing concentration of hydrochloric acid. The distribution coefficients of metal ions in water are larger than those in various hydrochloric acid concentration. The distribution coefficients of alkali and alkaline earth metal ions for the Dowex 50W-X8 ion exchange resin increased with decreasing hydrochloric acid concentration. Especially, the distribution coefficients of alkaline earth metal ions increased rapidly compared to those for alkali metal ions. The distribution coefficients of alkali and alkaline earth metal ions on HDBPDA ion exchange resin increased in a linear manner with decreasing acid concentration, and the slope, d log Kd/d log $M_{HCl}$ is about -0.2. Of the distribution cofficients of alkali metal ions on Dowex 50W-X8, at range of moderate hydrochloric acid concentration, the slope is about -1, while the slope for alkaline earth metal ions is about -2. However, at very low hydrochloric acid concentration, the linear variation between distribution coefficient and acid concentration was not occurred, but the slope was deviated from above values at low acid concentration.

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