• Title/Summary/Keyword: P-M interaction

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Preparation of High-Purity Urokinase Using Single-Step Hydrophobic Interaction Chromatography with p-Aminobenzamidine Ligand

  • Cao, Xue-Jun;Zhou, Jian-Hua;Huang, Zhen-Hui;Wu, Xing-Yan;Hur, Byung-Ki
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.196-203
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    • 2002
  • A novel process for urokinase purification was studied using p-aminobenzamidine as the ligand and sepharose 4B as the matrix. The adsorption, washing, and elution conditions were optimized by an unusual method. An adsorption buffer containing 2.5 M NaCl and $1\%$ Tween 80 facilitated the adsorption of urokinase on the affinity media and prevented contaminants from binding to the p-aminobenzamidine affinity gel. It was found that $5\%$ Tween 80 removed most of the contaminants from the affinity column. A 0.2 M glycine elution buffer containing 0.5 M NaCl (pH 3.0) was found to have a strong elution ability with a high recovery and purity of urokinase. A crude urokinase material of231 IU/mg protein from human urine was purified to 124,300 IU/mg protein with a purification factor of 538 and yield of $86.7\%$. As a result, a high purity urokinase was obtained with only a single affinity chromatography step. The purification process was successfully scaled-up to a 2-1 chromatography column. The resulting urokinase eluate could be directly lyophilized, thereby complying with Chinese pharmacopoeia (1995 version) standards.

Glutathione S-transferase T1, M1 and P1 Genetic Polymorphisms and Susceptibility to Colorectal Cancer in Turkey

  • Gorukmez, Ozlem;Yakut, Tahsin;Gorukmez, Orhan;Sag, Sebnem Ozemri;Topak, Ali;Sahinturk, Serdar;Kanat, Ozkan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3855-3859
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    • 2016
  • Colorectal cancer (CRC) is reproted to be the third most common cancer worldwide and the fourth most common cause of cancer related deaths. CRC is considered to be a multifactorial disease whose risk varies due to the complex interaction between individual genetic basis and disposure to multiple endogenous factors. Glutathione S-transferases are pro-carcinogenic in CRC and are required for the conjugation between chemotherapeutics and broad spectrum xenobiotics. One hundred and eleven patients with CRC and 128 control subjects without any cancer history were enrolled in this study. Multiplex PCR was applied to determine polymorphisms for the GSTT1 and M1 genes, and PCR-RFLP was applied for the GSTP1 (Ile105Val) gene polymorphism. Values p<0.05 were defined as statistically significant. We detected a significant high correlation between predisposition for CRC and presence of the Ile/Ile genotype of the GSTP1 (IIe105Val) gene polymorphism, but we did not find a significant relationship between predisposition for CRC and GSTT1 and M1 deletion polymorphisms. In addition, we did not determine a relationship between GSTT1, M1 and P1 gene polymorphisms and any clinicopathological features of CRC. GSTT1 null/GSTM1 positive and GSTT1 null/GSTM1 positive/GSTP1 Ile/Ile genotypes were significantly higher in the patient group. Our results revealed that there is no relationship among CRC, its clinicopathologic features, and GSTT1 M1 gene polymorphisms. However, there was a significant correlation between CRC and the GSTP1 Ile/Ile genotype. Further studies with larger patient groups are required to delineate the relationships between GST gene polymorphisms and the clinicopathologic features of CRC in Turkey.

A Study of Characteristics of lnxGa1-xP by Photoreflectance measurement (Photoreflectance 측정에 의한 InxGa1-xP의 특성 연구)

  • Kim D. L.;Yu J. I.
    • Laser Solutions
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    • v.8 no.3
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    • pp.5-10
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    • 2005
  • [ $InxGa_{1-x}P/GaAs$ ] structures were grown by chemical beam epitaxy(CBE), Pure phosphine($PH_3$) gases were used as group V sources. for the group III sources, TEGa, TmIn were used. $InxGa_{1-x}P$ epilayer was grown on SI-GaAs substrate and has a 1-${\mu}m$ thick. We have investigated the characteristics of $InxGa_{1-x}P$ by the photoreflectance(PR) spectroscopy, The PR spectrum of $InxGa_{1-x}P$ shows third-derivative feature whose Peaks Provide energy gap. The energy gap of $InxGa_{1-x}P$ has deduced composition x. From temperature dependance of PR spectra, temperature coefficient is $dEg/dT=-3.773{\times}10^{-4}$ eV/K, and Varshni coefficients $\alpha$ and $\beta$ values obtained $4{\times}10^4$ eV/K and 267 K respectively. Also, interaction $\alpha$B was 19.4 meV using the Bose-Einstein temperature relation, and $\Theta$ value related the average phonon frequency were 101.4 K. In particular, shoulder peak related to defects observed in PR signal that measured in temperature 82 K.

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Modeling and Parametric Studies on Moment-Curvature Relation of a Reinforced Concrete Column Subject In Axial-toad and Bi-Axil Moment (축하중과 이축모멘트를 받는 철근콘크리트 기둥의 모멘트-곡률에 관한 모델링 및 변수고찰)

  • 이차돈;최기봉;차준실;김성진
    • Journal of the Korea Concrete Institute
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    • v.14 no.5
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    • pp.677-688
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    • 2002
  • A analytical model is developed which can simulate a complete inelastic biaxial moment-curvature relations of a reinforced concrete column. The model can simulate sudden drop in moment capacity after peak moment and due to spalling of cover concrete. Parametric studies are performed examine the effects of constituent material properties as well as topological arrangement of reinforcements on moment-curvature relations and P-M interaction curve. It has been analytically observed that ductility of a reinforced concrete column is influenced mostly by magnitude of the axial load and spacings or the volume of lateral reinforcements. Compared to ACI P-M interaction curve, overall increase about 10% in square root of sum of squares of axial force and moment, and about 20% in peak load are observed for the columns reinforced according to ACI seismic design code.

Development of ELISA System for Screening of Specific Binding Inhibitors for Src Homology (SH)2 Domain and Phosphotyrosine Interactions

  • Lee, Sang-Seop;Lee, Kyung-Im;Yoo, Ji-Yun;Jeong, Moon-Jin;Park, Young-Mee;Kwon, Byoung-Mog;Bae, Yun-Soo;Han, Mi-Young
    • BMB Reports
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    • v.34 no.6
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    • pp.537-543
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    • 2001
  • In the present study, an in vitro ELISA system to assess the interaction between Src homology (SH)2 domains and phosphotyrosine that contain peptides was established using purified GST-conjugated SH2 proteins and synthetic biotinylated phosphotyrosine that contain oligopeptides. The SH2 domains bound the relevant phosphopeptides that were immobilized in the streptavidin-coated microtiter plate in a highly specific and dose-dependent manner. The epidermal growth factor receptor (EGFR)-, T antigen (T Ag)-, and platelet-derived growth factor receptor (PDGFR)-derived phosphopeptides interacted with the growth factor receptor binding protein (Grb)2/SH2, Lck/SH2, and phosphatidyl inositol 3-kinase (PI3K) p85/SH2, respectively. No cross-reactions were observed. Competitive inhibition experiments showed that a short phosphopeptide of only four amino acids was long enough to determine the binding specificity. Optimal concentrations of the GST-SH2 fusion protein and phosphopeptide in this new ELISA system for screening the binding blockers were chosen at 2nM and 500nM, respectively. When two candidate compounds were tested in our ELISA system, they specifically inhibited the Lck/SH2 and/or p85/SH2 binding to the relevant phosphopeptides. Our results indicate that this ELISA system could be used as an easy screening method for the discovery of specific binding blockers of protein-protein interactions via SH2 domains.

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Effect of bond slip on the performance of FRP reinforced concrete columns under eccentric loading

  • Zhu, Chunyang;Sun, Li;Wang, Ke;Yuan, Yue;Wei, Minghai
    • Computers and Concrete
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    • v.24 no.1
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    • pp.73-83
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    • 2019
  • Concrete reinforced with fiber reinforced polymer (FRP) bars (FRP-RC) has attracted a significant amount of research attention in the last three decades. A limited number of studies, however, have investigated the effect of bond slip on the performance of FRP-RC columns under eccentric loading. Based on previous experimental study, a finite-element model of eccentrically loaded FRP-RC columns was established in this study. The bondslip behavior was modeled by inserting spring elements between FRP bars and concrete. The improved Bertero-Popov-Eligehausen (BPE) bond slip model with the results of existing FRP-RC pullout tests was introduced. The effect of bond slip on the entire compression-bending process of FRP-RC columns was investigated parametrically. The results show that the initial stiffness of bond slip is the most sensitive parameter affecting the compression-bending performance of columns. The peak bond stress and the corresponding peak slip produce a small effect on the maximum loading capacity of columns. The bondslip softening has little effect on the compression-bending performance of columns. The sectional analysis revealed that, as the load eccentricity and the FRP bar diameter increase, the reducing effect of bond slip on the flexural capacity becomes more obvious. With regard to bond slip, the axial-force-bending-moment (P-M) interaction diagrams of columns with different FRP bar diameters show consistent trends. It can be concluded from this study that for columns reinforced with large diameter FRP bars, the flexural capacity of columns at low axial load levels will be seriously overestimated if the bond slip is not considered.

The antinociceptive effect of artemisinin on the inflammatory pain and role of GABAergic and opioidergic systems

  • Dehkordi, Faraz Mahdian;Kaboutari, Jahangir;Zendehdel, Morteza;Javdani, Moosa
    • The Korean Journal of Pain
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    • v.32 no.3
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    • pp.160-167
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    • 2019
  • Background: Pain is a complex mechanism which involves different systems, including the opioidergic and GABAergic systems. Due to the side effects of chemical analgesic agents, attention toward natural agents have been increased. Artemisinin is an herbal compound with widespread modern and traditional therapeutic indications, which its interaction with the GABAergic system and antinoniceptive effects on neuropathic pain have shown. Therefore, this study was designed to evaluate the antinociceptive effects of artemisinin during inflammatory pain and interaction with the GABAergic and opioidergic systems by using a writhing response test. Methods: On the whole, 198 adult male albino mice were used in 4 experiments, including 9 groups (n = 6) each with three replicates, by intraperitoneal (i.p.) administration of artemisinin (2.5, 5, and 10 mg/kg), naloxone (2 mg/kg), bicuculline (2 mg/kg), saclofen (2 mg/kg), indomethacin (5 mg/kg), and ethanol (10 mL/kg). Writhing test responses were induced by i.p. injection of 10 mL/kg of 0.6% acetic acid, and the percentage of writhing inhibition was recorded. Results: Results showed significant dose dependent anti-nociceptive effects from artemisinin which, at a 10 mg/kg dose, was statistically similar to indomethacin. Neither saclofen nor naloxone had antinociceptive effects and did not antagonize antinociceptive effects of artemisinin, whereas bicuculline significantly inhibited the antinocicptive effect of artemisinin. Conclusions: It seems that antinocicptive effects of artemisinin are mediated by $GABA_A$ receptors.

Purification and Characterization of β-Xylosidase from Paenibacillus sp. DG-22 (Paenibacillus sp. DG-22로부터 β-xylosidase의 정제 및 특성분석)

  • Lee, Tae-Hyeong;Lim, Pyung-Ok;Lee, Yong-Eok
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1341-1346
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    • 2007
  • An intracellular ${\beta}-xylosidase$ from Paenibacillus sp. DG-22 was purified to homogeneity by ion-exchange, hydrophobic interaction and gel-filtration chromatography. The molecular weight of the enzyme was measured to be 156,000 by gel filtration and 80,000 by SDS-PAGE, indicating that the enzyme consisted of two identical subunits. The purified enzyme exhibited maximum activity at $65^{\circ}C$ and pH 5.5. It retained 89% of its initial activity up to 60 min at $60^{\circ}C$ and had a half-life of 25 min at $65^{\circ}C$. The enzyme was highly specific for pNPX as the substrate. It showed little or no activity against other p-nitrophenyl glycosides and xylans. The $K_m\;and\;V_{max}$ for pNPX was 0.53 mM and 3.18 U/mg protein, respectively. The ${\beta}-xylosidase$ was strongly inhibited by $Ag^+,\;Fe^{2+},\;Hg^{2+}\;and\;Zn^{2+}$ and slightly activated by DTT. The hydrolysis product from xylobiose, xylotriose, and xylotetraose was xylose.

Interaction between norepinephrine and angiotensin II on Na+ uptake in primary cultured rabbit renal proximal tubule cells (초대배양한 토끼 신장 근위세뇨관세포의 Na+ uptake에 대한 norepinephrine 과 angiotensin II의 상호작용)

  • Park, Soo-hyun;Koh, Hyun-ju;Han, Ho-jae
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.525-534
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    • 1998
  • 신장 근위세뇨관세포들은 사구체에서 여과된 물질의 재흡수, 분비 및 대사에 관여하는 여러 호르몬들의 수용체들을 가지고 있다. 이들중에서 norepinephrine(NE)과 angiotensin II(ANG II)는 $Na^{+}/H^+$ 상호운반계를 조절함으로써 혈압조절에 관여하는 것으로 알려져 있으나 이들의 상호관계에 대해선 연구보고가 많지 않다. 본 연구는 초대배양한 토끼신장 근위세뇨관세포를 이용한 $Na^+$ uptake 실험을 통하여 NE이 어떠한 수용체를 통하여 $Na^{+}/H^+$ 상호운반계를 조절하는지 그리고 이러한 작용에 있어서 NE과 ANG II의 상호관계를 알아보고자 실시하였다. NE(>$10^{-9}M$)은 $Na^+$ uptake를 유의성 있게 증가시켰다($10^{-9}M$ NE : $27{\pm}4%$ increase vs. Control;p < 0.05). $\alpha$ 길항제(phentolamine, $10^{-10}M$)는 NE($10^{-9}M$)에 의해 유도된 $Na^+$ uptake를 유의성 있게 차단하였으나 (phentolamine+NE : $29{\pm}5%$ inhibition vs. NE ; p〈 0.05), ${\alpha}_1$ (pra-zosin, $10^{-10}M$) 및 ${\alpha}_2$ 길항제(yohimbine, $10^{-10}M$)는 부분적으로 차단하였다. ${\beta}$ 길항제(propra-nolol, $10^{-10}M$)도 역시 NE에 의해 유도된 $Na^+$ uptake를 유의성 있게 차단하였으나(propranolol+NE : $24{\pm}6%$ inhibition vs. NE ; p< 0.05), ${\beta}_1$(atenolol, $10^{-10}M$) 및 ${\beta}_2$ 길항제(butoxamine, $10^{-10}M$)는 부분적으로 차단하였다. 이러한 결과들은 NE에 의해 유도된 $Na^+$ uptake 증가작용은 ${\alpha}$(${\alpha}_1$${\alpha}_2$ )와 ${\beta}$(${\beta}_1$${\beta}_2$) 수용체 모두를 통하여 일어난다는 것을 시사해주고 있다. ANG II($10^{-11}M$) 또는 NE(${\alpha}_1$, ${\alpha}_2$, ${\beta}_1$, ${\beta}_2$ 작동제) 단독처리군의 $Na^+$ uptake는 대조군에 비해 유의성 있게 증가하였으나 (ANG II : $23{\pm}9%$ increase vs. Control; p < 0.05), 병합처리시 상승작용은 나타나지 않았다. ${\alpha}$ 또는 ${\beta}$ 길항제 처리시 NE 및 ANG II에 의해 유도되었던 $Na^+$ uptake 증가는 유의성 있게 차단되었다(phentolamine+NE+ANG II : $25{\pm}3%$ inhibition, propranolol+NE+ANG II : $24{\pm}6%$ inhibition vs. NE+ANG II, respectively ; p〈 0.05). 이 결과들은 $Na^+$ uptake에 있어서 ${\alpha}$(${\alpha}_1$${\alpha}_2$)와 ${\beta}$(${\beta}_1$${\beta}_2$) 수용체와 ANG II의 관련성을 시사해 준다. 결론적으로 토끼 신장 근위세뇨관세포에서 NE은 ${\alpha}_1$, ${\alpha}_2$, ${\beta}_1$${\beta}_2$ 수용체를 통하여 $Na^+$+ uptake를 증가시켰으며 이들 수용체는 ANG II $Na^+$ uptake 증가작용에 관여하였다.

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Study on the Solubility of U(VI) Hydrolysis Products by Using a Laser-Induced Breakdown Detection Technique (레이저유도파열검출 기술을 이용한 우라늄(VI) 가수분해물의 용해도 측정)

  • Cho, Hye-Ryun;Park, Kyoung-Kyun;Jung, Euo-Chang;Jee, Kwang-Yong
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.5 no.3
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    • pp.189-197
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    • 2007
  • The solubility of U(VI) hydrolysis products was determined by using a laser-induced breakdown detection (LIBD) technique. The experiments were carried out at uranium concentrations in range from $2{\times}10^{-4}\;M\;to\;4{\times}10^{-6}\;M$, pH values between 3.8 and 7.0, the constant ionic strength of 0.1 M $NaClO_4$ and the temperature of $25.0{\pm}0.1^{\circ}C$. The solubility product of U(VI) hydrolysis products was calculated from LIBD results by using the hydrolysis constants selected in NEA-TDB. The solubility product extrapolated to zero ionic strength, ${\log}K^{\circ}_{sp}=-22.85{\pm}0.23$ was calculated by using a specific ion interaction theory (SIT). The spectral features of ionic species in uranium solutions were investigated by using a conventional UV-visible absorption spectrophotometer and a fluorophotometer, respectively, $(UO_2)_2(OH)_2^{2+}\;and\;(UO_2)_3(OH)_5^+$ were dominant species at uranium concentration of $2{\times}10^{-4}\;M$.

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