• Title/Summary/Keyword: inhibition kinetics

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Distribution of Insulin-Like Growth Factor-Binding Proteins(IGFBPs) and IGFBP-3 Proteolysis in Noninsulin-Dependent Diabetes Mellitus Serum (인슐린 비의존형 당뇨병 환자의 혈청 중 Insulin-Like Growth Factor-Binding Proteins(IGFBPs)의 분포 및 IGFBP-3의 분해)

  • Lee, Hwa-Jin;Kim, Sung-Hyun;Kwon, Mi-Jin;Nam, Taek-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.285-290
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    • 1997
  • The insulin-like growth factors(IGFs) are bound to several binding proteins(IGFBPs) that appear to regulate IGF transport, receptor binding, and its action. The concentration of these peptides are altered by catabolic conditions. To determine IGF-I and IGFBP levels in noninsulin-dependent diabetes mellitus (NIDDM), sera was obtained from 5 patients and 7 controls. Serum levels of IGF-I in NIDDM were lower than those in either of the controls. By western immunoblot analysis, especially IGFBP-1 levels are increased, whereas IGFBP-3 levels decreased and their fragments was increased in NIDDM serum. IGFBP-3 proteolytic activity in NIDDM sera was inhibited by phenylmethylsulfonylfluoride (PMSF), aprotinin, and ethylenediaminetetraacetic acid(EDTA). This pattern of inhibition was consistent with a metal-dependent serine protease. By gelatin zymography, these proteolytic enzymes were identified as the size of 97 and 69 kDa. IGFBP-1, which is primarily insulin regulated, was increased in NIDDM and may modulate circulating IGF-I levels by regulating capillary passage of IGF-I. IGFBP-3 proteolysis markedly reduces its affinity for the IGFs, particularly for IGF-I. This accelerates their kinetics of dissociation, thereby increasing the proportions of IGF-I in free form and its availability to the cells.

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Effect of chloride ions on the catalytic properties of human pancreatic α-amylase isozyme produced in Pichia pastoris (Pichia pastoris에서 생산된 인체 췌장 α-아밀레이스 동질효소의 촉매활성에 대한 염소이온의 영향)

  • Kim, Min-Gyu;Kim, Young-Wan
    • Korean Journal of Food Science and Technology
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    • v.48 no.4
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    • pp.341-346
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    • 2016
  • The AMY2B gene, encoding human pancreatic ${\alpha}$-amylase isozyme (HPA II), was expressed in Pichia pastoris, and the effects of chloride ions on HPA II activity toward starch substrates were investigated. As seen with chloride ion-dependent ${\alpha}$-amylases-including HPA I, the isozyme of HPA II-chloride ions increased enzyme activity and shifted the optimal pH to an alkaline pH. The activity enhancement by chloride was more significant at pH 8 than that at pH 6, suggesting that the protonation state of the general acid/base catalyst of HPA II was important for the hydrolysis of starches at an alkaline pH because of the increase in its $pK_a$ by chloride ions. The turnover values for cereal starches as the substrates markedly increased in the presence of chloride by up to 7.2-fold, whereas that for soluble starch increased by only 1.7-fold. Chloride inhibited substrate hydrolysis at high substrate concentrations, with $K_i$ values ranging from 6 to 15 mg/mL.

Determination of the Optimum NH$_3$-N/NO$_2$-N Ratio by Anaerobic Batch Test in Anaerobic Ammonium Oxidation Process (혐기성 암모늄 산화공정에서 혐기성 회분식 실험에 의한 NH$_3$-N/NO$_2$-N의 최적비 산정)

  • Lee, Hwan-Hee;Kim, I-Jung;Jung, Jin-Young;Kim, Jee-Hyung
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.7
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    • pp.700-704
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    • 2008
  • Nitrite and free ammonia have been known as substrate inhibitors in anaerobic ammonium oxidation. To reduce inhibitory effect of these substrates, the NH$_3$-N/NO$_2$-N ratio in the influent could be properly controlled in anaerobic ammonium oxidation process. Five kinds of NH$_3$-N/NO$_2$-N ratio were assayed in batch to find optimum NH$_3$-N/NO$_2$-N ratio, curtailing substrate inhibition. As the results of batch test, the highest T-N removal efficiency of 88% was obtained at 1.00 : 1.30 of NH$_3$-N/NO$_2$-N ratio. In addition, rate constants for ammonium and nitrite in zero-order kinetics were found to be the highest values as 7.66 mg/L$\cdot$hr and 11.89 mg/L$\cdot$hr at 1.00 : 1.30 ratio, respectively. However, as for the specific anammox activity, the ratio of NH$_3$-N/NO$_2$-N ratio was recommended as 1 : 1.15 which can maintain the highest SAA during continuous operation and preclude the accumulation of nitrite in the reactor.

Acidic pH-activated $Cl^-$ Current and Intracellular $Ca^{2+}$ Response in Human Keratinocytes

  • Park, Su-Jung;Choi, Won-Woo;Kwon, Oh-Sang;Chung, Jin-Ho;Eun, Hee-Chul;Earm, Young-E;Kim, Sung-Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.4
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    • pp.177-183
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    • 2008
  • The layers of keratinocytes form an acid mantle on the surface of the skin. Herein, we investigated the effects of acidic pH on the membrane current and $[Ca^{2+}]_c$ of human primary keratinocytes from foreskins and human keratinocyte cell line (HaCaT). Acidic extracellular pH ($pH_e{\leq}5.5$) activated outwardly rectifying $Cl^-$ current ($I_{Cl,pH}$) with slow kinetics of voltage-dependent activation. $I_{Cl,pH}$ was potently inhibited by an anion channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 73.5% inhibition at 1${\mu}$M). $I_{Cl,pH}$ became more sensitive to $pH_e$ by raising temperature from $24^{circ}C$ to $37^{circ}C$. HaCaT cells also expressed $Ca^{2+}$-activated $Cl^-$ current ($I_{Cl,Ca}$), and the amplitude of $I_{Cl,Ca}$ was increased by relatively weak acidic $pH_e$ (7.0 and 6.8). Interestingly, the acidic $pH_e$ (5.0) also induced a sharp increase in the intracellular [$Ca^{2+}$] (${\triangle}[Ca^{2+}]_{acid}$) of HaCaT cells. The ${\triangle}[Ca^{2+}]_{acid}$ was independent of extracellular $Ca^{2+}$, and was abolished by the pretreatment with PLC inhibitor, U73122. In primary human keratinocytes, 5 out of 28 tested cells showed ${\triangle}[Ca^{2+}]_{acid}$. In summary, we found $I_{Cl,pH}$ and ${\triangle}[Ca^{2+}]_{acid}$ in human keratinocytes, and these ionic signals might have implication in pathophysiological responses and differentiation of epidermal keratinocytes.

N-(4-[$^{18}F$]Fluoromethylbenzyl)spiperone : A Selective Radiotracer for In Vivo Studies of Dopamine $D_2$ Receptors (N-(4-[$^{18}F$Fluoromethylbenzyl)spiperone : 유력한 도파민 $D_2$ 수용체 선택성 방사성리간드)

  • Kim, Sang-Eun;Choe, Yearn-Seong;Chi, Dae-Yoon;Lee, Kyung-Han;Choi, Yong;Kim, Byung-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.31 no.4
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    • pp.421-426
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    • 1997
  • We evaluated the in vivo kinetics, distribution, and pharmacology of N-(4-[$^{18}F$]fluoromethylbenzyl)spiperone ([$^{18}F$]FMBS), a newly developed derivative of spiperone, as a potentially more selective radiotracer for the dopamine (DA) $D_2$ receptors. Mice received 1.9-3.7 MBq (1.8-3.6 nmol/kg) of [$^{18}F$]FMBS by tail vein injection. The time course and regional distribution of the tracer in brain were assessed. Blocking studies were carried out by intravenously preinjecting DA $D_2$ receptor blockers (spiperone, butaclamol) as well as drugs with high affinity for DA $D_1$ (SCH 23390), DA transporter (GBR 12909), and serotonin $S_2$ ($5-HT_2$) (ketanserin) sites. After injection of the tracer, the radioactivity in striatum increased steadily over time, resulting in a striatal-to-cerebellar ratio of 4.8 at 120 min postinjection. By contrast, the radioactivity in cerebellum, frontal cortex, and remaining cortex washed out rapidly. Preinjection of unlabeled FMBS (1 mg/kg) and spiperone (1 mg/kg) reduced [$^{18}F$]FMBS striatal-to-cerebellar ratio by 41% and 80%, respectively. (+)-Butaclamol (1 mg/kg) blocked 80% of the striatal [$^{18}F$]FMBS binding, while (-)-butaclamol (1 mg/kg) did not. Preinjection of SCH 23390 (1 mg/kg) and GBR 12909 (5 mg/kg) had no significant effect on [$^{18}F$]FMBS binding. Ketanserin (1 mg/kg), a ligand for the $5-HT_2$ receptors, did not cause significant inhibition either in striatum, in frontal cortex, or the remaining cortex. The results demonstrate that [$^{18}F$]FMBS labels DA $D_2$ receptors selectively in vivo in the mouse brain. It may hold promise as a selective radiotracer for studying DA $D_2$ receptors in vivo by PET.

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The effect of HRT, current density, and packing ratio on nitrate nitrogen removal efficiency and current efficiency in BRM-BER (고정상 담체를 충진한 BER에서 HRT, 전류밀도 및 담체 충진율 변화가 질산성 질소 제거효율과 전류이용효율에 미치는 영향)

  • Whang, Gye-Dae;Lee, Sang-Keun;Sung, Hae-Chang
    • Journal of Korean Society of Water and Wastewater
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    • v.24 no.4
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    • pp.433-442
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    • 2010
  • BER at different packing ratios of bio-ring media(BRM) was tested to investigate the effect of varying hydraulic retention time (HRT) and current density on the nitrate removal and current efficiency. In the preliminary batch mode experiment of BERs, current density was applied at 2.0 A/$m^2$, 4.0 A/$m^2$, 4.8 A/$m^2$, which correspond to the designation of reactor #1, #2, #3, respectively. The reactor #2 showed a highest nitrate removal rate of 162.0 mg $NO_3{^-}$-N/L/d, and the kinetics of nitrate removal rate was defined as the Zero order reaction. In the primary experiment of BERs, four BERs packed with BRM were operated in varying HRT and current, and the packing ratios of reactor #1, #2, #3 and #4 were 0%, 8%, 16%, 24%. respectively. This results of the experiments indicated that the nitrate removal rate and current efficiency were increased significantly cause of growing of autotrophic denitrification microorganisms on the surface of cathode and media by increasing of the current density and decreasing of HRT. However, The decreasing of nitrate removal rate and current efficiencies were observed in the condition of HRT of 5.25 hr and 4.8 A/$m^2$ of current density. With more increasing current density and decreasing of HRT, the hydrogen inhibition occurred at the surface of cathode. Moreover, nitrate removal rate by autotrophic denitrification microorganisms attached on the media surface was observed to be limited by no longer increasing dissolved hydrogen concentration of each reactor. In conclusion, the highest nitrate nitrogen removal and current efficiency could be achieved when the BER was operated at the conditions of 7 hr HRT, current density of 4.0 A/$m^2$, and 16% packing ratio. And it was found that the amount of nitrate removal by microorganisms attached on the surface of cathode and media (BRM) was 178.2 mg/L and 52.2 mg/L respectively. and the amount of nitrate removal per MLVSS was 0.435 g $NO_3{^-}$-N/g $MLVSS{\cdot}d$ and 0.336 $NO_3{^-}$-N/g $MLVSS{\cdot}d$.

Effects of the immobilization of heparin and rhPDGF-BB to titanium surfaces for the enhancement of osteoblastic functions and anti-inflammation

  • Huh, Jung-Bo;Lee, Jeong-Yo;Lee, Kyung-Lae;Kim, Sung-Eun;Yun, Mi-Jung;Shim, Ji-Suk;Shim, June-Sung;Shin, Sang-Wan
    • The Journal of Advanced Prosthodontics
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    • v.3 no.3
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    • pp.152-160
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    • 2011
  • PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

Inhibitory Action of YJA20379, a New Proton Pump Inhibitor on Helicobacter Pylori Growth and Urease

  • Woo, Tae-Wook;Chang, Man-Sik;Chung, Young-Kuk;Kim, Kyu-Bong;Sohn, Sang-Kwon;Kim, Sung-Gyu;Choi, Wahn-Soo
    • Archives of Pharmacal Research
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    • v.21 no.1
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    • pp.6-11
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    • 1998
  • The activities of two types of antiulcer agents against 9 strains of Helicobacter pylori (H. pylori) were determined by the agar dilution method. The antiulcer agents were YJA20379, a newly synthesized proton pump inhibitor developed by Yung-jin Pharmaceutical company, and omeprazole. Both compounds were found to have significant activities against this organism. The MIC values of YJA20379 and omeprazole were 11.7 and $31.25{\mu.g/ml}$ respectively. In addition, the inhibitory potency of both compounds was investigated on H. pylori urease which is believed to be an important colonization and virulence factor in the pathogenesis of gastritis and peptic ulcers. These compounds dose-dependently inhibited urease extracted with distilled water and their $IC_50$ values were $16.4{\times}10^{-5} M and 14.3{\times}10^{-5}M,$ respectively. In addition, a pH-dependent study to determine whether inhibitory potency would be activated by acid condition was performed. It was found that unlike omeprazole, YJA20379 was not affected by acid condition. To determine the inhibition pattern and optimal concentration of substrate, kinetics were evaluated at various pH levels (pH 5.0, 7.0, and 8.5). The data show that YJA20379 noncompetitively inhibited H. pylori urease and $K_M/K_i$values were 0.96 $mM/60{\mu}M (pH 5.0), 0.56 mM/141.5 {\mu}M (pH 7.0)$, and $1.94mM/34{\mu}M (pH 8.5)$, respectively. Based on data obtained, it is concluded that YJA20379 is a significant inhibitor of H. pylori growth and urease and therefore, taking these results into consideration, YJA20379 might be a beneficial therapy for gastritis and peptic ulcers induced by H. pylori.

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Studies on Immunomodulatory and Antioxidant Activities of Astragali membranacei Radix according to the Cultivated Years (황기(黃芪)의 재배 년수에 따른 면역 및 항산화 활성 연구)

  • Jung, Chul
    • Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
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    • v.1 no.1
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    • pp.53-90
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    • 2005
  • Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

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Analysis of the Factors Affecting Anaerobic Thermophilic Digestibility of Food Wastes (음식물쓰레기의 고온 혐기성 소화도에 미치는 요소에 대한 분석)

  • Kim, Do Hee;Hyun, Seung Hoon;Kim, Kyung Woong;Cho, Jaeweon;Kim, In S.
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.2
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    • pp.130-139
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    • 2000
  • Serial basic tests were conducted for the determination of fundamental kinetics and for the actual application of kinetic parameter to food waste digestion with precise measurement of methane production under a thermophilic condition. The effects of food particle size, sodium ion concentration, and volatile solid (VS) loading rate on the anaerobic thermophilic food waste digestion process were investigated. Results of serial test for the determination of fundamental kinetic coefficients showed the value of k (maximum substrate utilization rate coefficient) and KS (half-saturation coefficient) as $0.24hr^{-1}$ and $700mg/{\ell}$, respectively, for non-inhibiting organic loading range. No inhibition effect was shown until $5g/{\ell}$ of sodium ion concentration was applied to a serum bottle reactor. However, the volume of methane gas was decreased gradually when the concentrations of more than $5g/{\ell}$ of sodium ion applied. All sizes of food waste particle showed the same constants (A : 0.45) but the maximum substrate utilization rate constant ($k_{HA}$) was inversely proportional to particle size. As an average particle size increased from 1.02 mm to 2.14 mm, $k_{HA}$ decreased from $0.0033hr^{-1}$ to $0.0015hr^{-1}$. The result reveals that particle size is one of the most important factors in anaerobic food waste digestion. There was no inhibition effect of sodium ion when VS loading rate was $30g/{\ell}$. And maximum injection concentration of VS loading rate was determined about $40g/{\ell}$.

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