• 제목/요약/키워드: Inhibition kinetics

검색결과 154건 처리시간 0.029초

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

  • 이화진;김성현;권미진;남택정
    • 한국식품영양과학회지
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    • 제26권2호
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    • pp.285-290
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    • 1997
  • 이상의 결과를 요약하연, 혈당값은 정상대조군 78{\pm}6.20mg/dl, 인슐린 비의존형 당뇨병 환자 $184{\pm}18.94mg/dl$로 인슐린 비의존형 당뇨병 환자의 혈당값이 유의적으로 높았다. IGF-I의 수준은 정상대조군 $9.32{\pm}1.48{\mu}g/ml$, 인슐린 비의존형 당뇨병 환자 $4.19{\pm}0.84{\mu}g/ml$로 인슐린 비의존형 당뇨병 환자의 IGF-I 수준이 유의적으로 낮았다. 인슐린 비의존형 당뇨병 환자의 혈청 중 IGFBP-1, -2는 증가한 반면, IGFBP-3, -4, -5는 감소하였다. IGFBP-3의 분해에 관여하는 효소는 PMSF, aprotinin, EDTA에 의해 저해되어 metal-dependent serine proteases임을 확인하였고, 효소의 크기가 대략 97,66kDa 이었으며 효소활성이 인슐린 비의존형 당뇨병 환자에게서 큼을 확인하였다.

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

  • 김민규;김영완
    • 한국식품과학회지
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    • 제48권4호
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    • pp.341-346
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    • 2016
  • HPA는 식품으로 섭취되는 녹말을 분해하는데 있어서 매우 중요한 역할을 수행하는 효소이기 때문에 HPA 효소 활성의 억제는 당뇨와 비만과 같은 질환의 예방과 치료에 있어서 의미를 가진다. 따라서 HPA는 당뇨병 치료와 비만 예방을 위한 새로운 식 의약품 소재 개발을 위한 주요 타깃 효소 중 하나이며, 새로운 소재의 개발을 위해서는 HPA의 반응 메커니즘을 비롯하여 천연 기질 분해 특성에 대한 이해가 반드시 필요하다. 본 연구에서는 HPA의 동질효소 중 연구가 거의 진행되지 않은 HPA II에 대한 효소 특성화를 진행하고자 P. pastoris 시스템을 이용하여 재조합 HPA II를 생산하였으며, 녹말 분해와 관련된 효소적 특성을 분석하였다. HPA II는 10 mM NaCl까지 농도 의존적으로 효소활성이 증가하였으며, 최적 활성을 위한 pH는 0 mM NaCl 조건에서 pH 6.5이었으나 10 mM NaCl조건에서 pH 7.5로 이동하는 특성을 보였으며, 이는 HPA I을 포함하는 염소이온 의존형 아밀레이스가 나타내는 전형적인 특징이다. 염소이온 존재 시 최적 pH가 염기성 pH 영역으로 이동하는 것은 염소 이온과 효소의 결합에 의해 HPA II의 산/염기 촉매 잔기의 $pK_a$값이 커진다는 것을 의미하며, 염소이온을 첨가하였을 때 녹말에 대한 가수분해 활성의 증대 정도가 산성 pH 영역보다 염기성 pH 영역에서 두드러지게 나타났다는 점이 이를 뒷받침하였다. 반응속도론적 분석 결과에 따르면 염소이온 존재 시 효소활성의 증대는 대부분 전환수(turnover number)의 증대에 의한 것으로 나타났으며, 가용성 녹말 보다 곡류 녹말에 대한 전환수의 증대가 크게 나타났다. 염소이온은 활성의 증대뿐만 아니라 고농도의 기질 조건에서 기질에 의한 효소 활성 억제 양상을 심화시키는 것으로 나타났다. 결론적으로 HPA II의 특징은 HPA I과 거의 유사한 경향을 나타내었으며, 염소이온 첨가여부에 따른 HPA II의 가수분해활성 결과를 바탕으로 향 후 HPA에 대한 곡류 녹말 가수분해 활성 억제제 개발을 위한 연구를 추진할 계획이다.

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

  • 이환희;김이중;정진영;김지형
    • 대한환경공학회지
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    • 제30권7호
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    • pp.700-704
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    • 2008
  • 혐기성 암모니아 산화공정에서 nitrite는 저해인자로 잘 알려져 있고, 최근에는 유리 암모니아 역시 anammox bacteria에 저해 영향을 주는 것으로 보고되고 있다. 유입수의 암모니아와 아질산의 비율이 연속운전에서 효과적인 질소제거에 중요한 인자가 되며, 연속운전 반응기에서는 유리 암모니아와 아질산의 축적을 방지하기 위해 유입수의 NH$_3$-N/NO$_2$-N-N비를 조절할 필요가 있다. 이에 본 연구에서는 다섯 가지 종류의 NH$_3$-N/NO$_2$-N-N비를 회분식 실험을 통해 잔류 암모니아성 질소와 아질산성 질소의 농도를 최소화하는 비를 조사하였다. 회분식 실험 결과 실험 26시간 후에 1.00 : 1.30의 비에서 88%에 달하는 총질소 제거율이 나타났다. 그리고 혐기성 암모늄 산화 반응은 0차 반응을 나타내었고, 암모니아와 아질산의 반응 속도상수는 1.00 : 1.30의 비에서 각각 7.66 mg/L$\cdot$hr과 11.89 mg/L$\cdot$hr로 가장 높게 나타났다. 혐기성 암모늄 산화균 활성도를 측정해본 결과 1.00 : 1.15의 비에서 미생물의 활성도가 가장 우수한 것으로 나타났다. 회분식 실험의 결과를 통해, 이론적 반응비과 비슷한 1.00:1.30에서는 반응속도가 크고 총질소 제거율도 높은 반면 혐기성 암모늄 산화균은 이론적 반응비보다 다소 낮은 아질산 농도에서 안정하다는 것을 확인할 수 있었다.

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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    • 제12권4호
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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 : 유력한 도파민 $D_2$ 수용체 선택성 방사성리간드 (N-(4-[$^{18}F$]Fluoromethylbenzyl)spiperone : A Selective Radiotracer for In Vivo Studies of Dopamine $D_2$ Receptors)

  • 김상은;최연성;지대윤;이경한;최용;김병태
    • 대한핵의학회지
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    • 제31권4호
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    • pp.421-426
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    • 1997
  • [$^{18}F$]FMBS는 도파민 $D_2$ 수용체 방사성추적자로서 유망한 성질을 지니고 있다. [$^{18}F$]FMBS는 비교적 높은 특이결합/비특이결합 비를 제공하며, 그 체내결합은 도파민 $D_2$ 수용체에 대하여 높은 특이성과 선택성을 보인다. 도파빈 $D_2$ 수용체 측정을 위한 보다 적합한 PET 방사성추적자를 얻기 위해서는 [$^{18}F$]FMBS의 낮은 뇌섭취와 느린 역학을 개선할 수 있는 화학적 구조의 변형이 시도되어야 하며, [$^{18}F$]FMBS는 이러한 시도의 골격이 될 수 있을 것으로 믿는다.

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

  • 황규대;이상근;성해창
    • 상하수도학회지
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    • 제24권4호
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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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    • 제3권3호
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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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    • 제21권1호
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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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황기(黃芪)의 재배 년수에 따른 면역 및 항산화 활성 연구 (Studies on Immunomodulatory and Antioxidant Activities of Astragali membranacei Radix according to the Cultivated Years)

  • 정철
    • 대한한방피부미용학회지
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    • 제1권1호
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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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음식물쓰레기의 고온 혐기성 소화도에 미치는 요소에 대한 분석 (Analysis of the Factors Affecting Anaerobic Thermophilic Digestibility of Food Wastes)

  • 김도희;현승훈;김경웅;조재원;김인수
    • 유기물자원화
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    • 제8권2호
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    • pp.130-139
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    • 2000
  • 일련의 회분식실험을 통하여 음식폐기물의 동력학상수 및 메탄가스 발생량을 측정하였으며 음식폐기물의 고온혐기성 소화시 입자의 크기, 나트륨이온의 농도, 휘발성 고형물의 부하량이 따른 운전인자에 대한 영향을 살펴보았다. 기초 동력학 실험에서는 최대 기질 분해 속도 상수 k는 $0.24hr^{-1}$로 나타났으며 반 포화상수 Ks는 $700mg/{\ell}$ 로 나타났다. 나트륨 농도에 따른 혐기성소화 영향에 관한 실험은 나트륨 농도가 $5g/{\ell}$ 까지는 총 메탄가스 발생량에 영향을 미치지 않았으나 점차로 농도가 증가함에 따라 총 메탄가스 발생량이 감소하기 시작하였으며 나트륨 농도가 $20g/{\ell}$ 로 증가시켰을 경우 이론적인 발생량의 50% 만을 발생하였다. 음식폐기물을 크기별로 분쇄하여 혐기성 소화를 시킨 후 Valentini 모델에 적용시킨 결과 모든 경우에 있어서 가수분해 상수 값 A는 0.45를 나타내었으며 입자의 크기가 1.02 mm에서 2.14 mm로 높아짐에 따라 최대 기질 분해 속도 상수 $k_{HA}$$0.0033hr^{-1}$에서 $0.0015hr^{-1}$로 감소하였다. 이것으로 음식폐기물의 혐기성 소화에 있어서 입자 크기가 중요한 운전인자 중의 하나임을 증명할 수 있었으며 휘발성 고형분의 부하량에 관한 연구에서는 최대 주입 휘발성 고형물 부하량은 $40g/{\ell}$ 로 나타났으며, 유기물 부하가 낮은 경우에서는 나트륨 이온에 따른 영향이 휘발성 고형물 제거에 나타나지 않았으나 $40g/{\ell}$ 인 경우 약간의 차이가 있는 것으로 판명되었다.

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