891 resultados para HMG-CoA reductase inhibition
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The feeding rate of the copepod Mesocyclops notius on two species of cladocerans (Daphnia carinata and Ceriodaphnia cornuta) decreased with increasing environmental concentration of 64-122 mum colonial Microcystis spp. Rate of copepod feeding on Moina micrura was unaffected by the presence of Microcystis spp. Mesocyclops notius rarely preyed on Diaphanosoma brachyurum.
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Microcystins isolated from toxic cyanobacteria are potent inhibitors of protein phosphatases 1 and 2A (PP1 and PP2A). The inhibitory effects of three structural variants of microcystins (microcystin-LR, -YR, and -RR) on protein phosphatases isolated and purified from the liver and kidney of grass carp (Ctenopharyngodon idellus) were investigated using the P-32 radiometric assay. The relationships between percentage inhibition of protein phosphatase activity and microcystin levels followed a typical dose-dependent sigmoid curve. These results were compared to those obtained from mouse PP2A. The degree and pattern of inhibition of both fish and mouse protein phosphatases by microcystins were similar. Protein phosphatases in crude fish tissue homogenates showed similar inhibition patterns as purified fish PP2A toward microcystins. (C) 2000 by John Wiley & Sons, Inc.
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A settlement inhibition assay using barnacle cyprid larvae, Balanus amphitrite, was developed with Cd2+ and phenol as standard reference toxicants. Mean percentage settlement of cyprid larvae showed a progressive reduction with increasing concentrations of Cd2+ and phenol. A significant reduction in settlement was found when cyprids were exposed to 0.1 mgL(-1) Cd2+ or 10 mgL(-1) phenol. The assay was used to assess the sublethal toxicity of three oil dispersants (Vecom B-1425 GL, Norchem OSD-570 and Corexit 9905) commonly used in Hong Kong waters. Results of this investigation show that the barnacle settlement inhibition assay can be incorporated into the battery of tests currently available for ecotoxicological assessment of marine contaminants. (C) 1997 Elsevier Science Ltd.
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The effects of sublethal concentrations of phenol and cadmium on the phototactic responses of the stage II nauplii of the barnacle Balanus amphitrite were investigated. Increased toxicant concentrations caused a reduction in phototactic responses. Balanus amphitrite nauplii exposed to nominal phenol concentrations of 100 ppm and higher for 1-12 h failed to exhibit phototactic responses, while longer exposure times of 24 and 48 h reduced the lowest observable effect concentration (LOECs) to 80 and 60 ppm, respectively. For cadmium, the LOECs, based on nominal concentrations, for B. amphitrite following 1, 6, 12, 24, and 48 h exposures were 20, 4.5, 4.0, 1, and 0.75 ppm, respectively. The LOECs can be significantly reduced by increasing the duration of exposure to the toxicants. A good relationship exists between the phototactic response and toxicant concentration as well as exposure time. Results of this study indicate that the toxicant-induced reduction in phototactic responses of barnacle larvae can be used in a sensitive, rapid screening test for ecotoxicological assessments. (C) 1997 by John Wiley & Sons, Inc.
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The corrosion inhibition behavior of benzotriazole, Na3PO4 and their mixture on carbon steel in 20 wt.% (0.628 mol l(-1)) tetra-n-butylammonium bromide aerated aqueous solution was investigated by weight-loss test, potentiodynamic polarization measurement, electrochemical impedance spectroscopy and scanning electron microscope/energy dispersive X-ray techniques. The inhibition action of BTA or SP or inhibitors mixture on the corrosion of carbon steel is mainly due to the inhibition of anodic process of corrosion. The results revealed that inhibitors mixtures have shown synergistic effects at lower concentration of inhibitors. At 2 g l(-1) BTA and 2 g l(-1) SP showed optimum enhanced inhibition compared with their individual effects.
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近二十多年来,基于对臭氧层衰减、紫外线B(UV-B)增强的担心,研究者希望了解到紫外线辐射对不同作物的影响情况,增强UV-B辐射条件下是否对作物的生长发育、产量质量构成威胁。在本试验中,我们首先探讨了双子叶作物黄瓜(Cucumis sativus)和大豆(Glycine max)对不同紫外波段的生物效应[分别为B-UVA(315-400 nm),N-UVA(315-340 nm),B-UVB(275-400 nm)和N-UVB(290-340 nm),UV-(>400nm)作对照]。我们观察到所有的UV波段处理都使黄瓜和大豆的生长受到抑制,并且细胞受到不同程度的氧化伤害;UV波段处理的作用效果与不同波段的紫外有效生物辐射剂量有关。处理差异在UV-B波段内部和UV-A波段内部同样存在。植物生长UV辐射公式(BSWF)能很好的预测本试验UV-B波段内的平均植物效应,但不能预测UV-A波段的植物效应。短波UV-A的生物作用强于长波UV-A。光合色素的变化与UV波谱差异和种间差异有关。在高的紫外/可见光背景下,UV-A处理同UV-B同样导致光合色素的降低,但黄瓜类胡萝卜素/叶绿素比例升高。与其他研究者的试验结果比较后,我们认为紫外线B辐射的生物效应一致性很高,但紫外线A波段的生物学效应存在较大争议。因此我们在本试验的基础上仅进行荞麦[苦荞(Fagopyrum tataricum Gaertn.)和甜荞(Fagopyrum esculentum Moench.)]对紫外线B波段的响应研究。 我们对苦荞品种-圆籽荞进行了连续两个生长季节的大田半控制试验以观察UV-B辐射对苦荞生长、发育、产量及叶片色素的影响;试验小区进行降低UV-B、近充足UV-B和增强UV-B辐射处理。我们的试验表明在不同强度UV-B辐射下苦荞的生长、地上部生物量积累及最终产量都有所下降,但苦荞的发育加快;当前条件下的日光紫外线B辐射对植物生长和产量也造成负面影响。植物光合色素被日光及增强UV-B辐射降低;UV化合物及卢丁含量在中低剂量的UV-B辐射强度下显著升高,但在高剂量的增强UV-B辐射下短期升高后迅速下降。我们的试验表明苦荞是一个对UV-B高度敏感的作物。苦荞对UV-B的敏感性与UV-B剂量、外界环境因素及生长季节有关。 单个苦荞品种的试验结果使我们认识到外界UV-B辐射已经对苦荞生长发育构成逆境条件,未来全球气候变化条件下增强紫外线B辐射可能使其处于更不利的生长环境中。因此我们有进行了多个种群进行UV-B响应观察并筛选耐性种群。我们对15个苦荞种群进行增强UV-B辐射处理(6.30 kJ m2 UV-BBE,模拟当地25%的臭氧衰减),我们观察苦荞UV-B辐射效应存在显著的种内差异,UV-B辐射对多数种群具有抑制作用,但对一些种群还有刺激作用。我们采用主成分分析方法与作物UV-B响应指数(RI)来评价苦荞作物UV-B辐射耐性。我们发现作物的UV-B耐性不仅与其原产地背景UV-B强度有关,而且与作物相对生长效率、次生代谢产物含量(如卢丁)及其他因素有关。我们观察到苦荞伸展叶总叶绿素变化与UV-B耐性成正相关;室内苦荞幼苗的UV-B辐射致死试验表明:苦荞种群死亡率与其UV-B耐性成负相关。 此外,我们对甜荞的UV-B辐射响应也进行了初步研究。选取美姑甜荞、巧家甜荞和云龙甜荞进行5个梯度的增强UV-B辐射室外模拟试验。我们观察到UV-B辐射显著降低了甜荞的生长、生物量及产量;并严重影响了甜荞的生殖生长,降低了花序数、种子数和结实率;并且UV-B辐射对甜荞的抑制作用存在显著的剂量效应。三种甜荞品种存在显著的种内差异,其中美姑品种UV-B耐性最强,且膜脂受UV-B辐射氧化伤害最小,这与该品种UV-B辐射下较高的GR酶活性、APX酶活性和PPO酶活性、以及含量更高的抗坏血酸有关。甜荞的次生代谢也受到增强UV-B辐射的影响,其香豆酰类化合物在UV-B辐射下升高显著,而槲皮素含量也在高剂量UV-B辐射下有所增加;卢丁含量依赖UV-B辐射剂量而变化,中低剂量UV-B辐射下其卢丁含量逐渐升高,但在高剂量辐射下逐渐下降。 通过对生长在高海拔地区的荞麦作物(苦荞和甜荞)进行的室外研究,我们认识到作物不同品种存在很大的耐性差异,这就为UV-B耐性育种创造了有利条件。进一步加大荞麦种质资源筛选力度并深入认识荞麦抗性机理,在此基础上通过杂交或其他基因融合手段培育抗性品种,对高剂量UV-B辐射地区的荞麦产量的提高将起到重要推动作用,并使荞麦生产能有效应对未来全球气候变化条件下UV-B辐射可能升高的威胁。 During last few decades, due to concern of ozone layer depletion and enhancement of ultraviolet B radiation(UV-B, 280-315 nm), the agronomist want to know the responses of different crop species to UV-B. In the first experiment of our study, the effect of different UV band [B-UVA(315-400 nm), N-UVA(315-340 nm), B-UVB(275-400 nm), N-UVB(290-340 nm)and UV-(>400nm, as control)] on the cucumber(Cucumis sativus)and soybean(Glycine max)were investigated in growth room. Spectra-dependent differences in growth and oxidation indices existed within UV-A bands as well as UV-B bands. The general biological effects of different band were UV- < B-UVA< N-UVA<N-UVB<B-UVB. The plant growth biologically spectra weighting function(BSWF)matched well with average plant response in UV-B region, but not in UV-A region. Shorter UV-A wavelength imposed more negative impact than longer UV-A wavelength did in both species. The effect on photosynthetic pigment was related to different UV bands and different species. The photosynthetic pigment content was decreased by UV-A spectra as well as UV-B spectra. In comparison with the results of previous studies, we found that the wavelength-dependent biological effect of ultraviolet B radiation has high consistency, but the biological effect of ultraviolet-A radiation was inconsistent. We narrow our following study on the effect of ultraviolet B radiation on the buckwheat(tartary buckwheat and common buckwheat). The tartary buckwheat(Fagopyrum tataricum Gaertn.)cultivars Yuanziqiao was grown in the sheltered field plots for two consecutive seasons under reduced, near-ambient and two supplemental levels of UV-B radiation. The crop growth, photosynthetic pigments, total biomass, final seed yield and thousand-grain weight were decreased by near-ambient and enhanced UV-B radiation, while crop development was promoted by enhanced UV-B radiation. Leaf rutin concentration and UV-B absorbing compound was generally increased by UV-B with the exception of 8.50 kJ m-2 day-1 supplemental levels. Our results showed that tartary buckwheat is a potentially UV-B sensitive species. Study on one cultivars showed that ambient solar radiation had present a stress to tartary buckwheat. This makes it necessary to observe the UV-B response of many cultivars and screen tolerant cultivars. Fifteen populations of tartary buckwheat were experienced enhanced UV-B radiation simulating 25% depletion of the stratospheric ozone layer in Kunming region, and plant responses in growth, morphology and productivity were observed. Principal components analysis(PCA)was used to evaluate overall sensitivity of plant response to UV-B as well as response index. The different populations exhibited significant differences in responses to UV-B. The photosynthetic pigments of young seedlings were also affected significantly under field condition. On the other hand, the healthy seedlings of different populations were exposed to the high level of UV-B radiation in growth chambers to determine the plant lethality rate. The plant tolerance evaluated by multivariate analysis was positively related to total plant chlorophyll change, but negatively related to lethality rate. In other hand, the UV-B responses of the other important cultivated buckwheat species, common buckwheat(Fagopyrum esculentum Moench.), were also studied preliminarily. Three widespread cultivated variety(Meigu, Qiaojia and Yunlong cultivars)were provided with five level of enhanced UV-B radiation outdoors. We observed that the crop growth, development and production were significantly decreased, and reproductive production, like anthotaxy number, seed number and seed setting ratio, was also decreased. Dose-dependent inhibition effect caused by enhanced UV-B radiation also existed in common buckwheat. Significant intraspecific difference existed in those three cultivars. The Meigu cultivars with dwarfed growth and lower production have highest UV-B tolerance as well as lowest damage in cell membrane, this could be associated with profound enhancements of glutathione reductase(GR)activity, ascorbate peroxidase activity and polyphenol oxidase activity as well as higher ascorbic acid concentration. The secondary metabolism was also affected by UV-B radiation, with profound elevation of coumarin compound and moderate increase of quercetin concentration. Rutin concentration was peaked in 5kJ m-2 UV-B. The contrasting effect of UV-B radiation on different populations indicated that there existed abundant genetic resources for selecting tolerant populations of common and tartary buckwheat. Much effort needed be pose on screening of buckwheat germplasm and clarification of mechanism of buckwheat tolerance to UV-B. On this base the tolerant cultivars could be bred by hybridization and other gene transfusion method, this would help increase buckwheat yield in high ambient UV-B region and counteract the effect of possible enhanced UV-B radiation in future.
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维生素(Vitamin)又称维他命,为“万年青”产品,是维持人体生命健康必需的一类低分子有机化合物质。维生素对人体健康的作用人们研究很多, 维生素可以增强人体对感染的抵抗力,降低出生缺陷及降低癌症和心脏病发病率等,一旦缺乏,肌体代谢就会失去平衡,免疫力下降,各种疾病,病毒就会趁虚而入;而维生素对作物影响的研究却很少。目前为止,尚无对用维生素浸种的方法来研究外源维生素是否对小麦种子萌发及幼苗生长起调节作用的报道,且对其在小麦抗逆性方面影响的研究甚少,对盐的胁迫抗性研究尚未有人报道。小麦(Triticum aestivum L.)属于拒盐的淡土性作物。盐害不利于小麦生长,严重影响小麦的产量和品质。本研究采用4 种不同维生素VB1、VC、VB6、VPP,分别对供试小麦品种川育12(红皮)、川育16(白皮)小麦浸种后,在一般自然条件下和逆境(盐胁迫条件)下,进行试验。探讨在正常情况下与在不同盐浓度条件下,各维生素及盐浓度对小麦发芽及幼苗生长的影响,并且比较两种不同皮色的小麦在相同盐胁迫条件下的差异表现,同时研究维生素处理的特异性,且哪种维生素对盐害缓解作用最佳。研究结果表明:在无盐胁迫(自然)条件下,对用4 种不同维生素VB1、VC、VB6、VPP 浸种小麦川育12、川育16 后的种子萌发及幼苗生长(幼苗的根长、根重、苗高、苗鲜重)的研究结果表明:4 种外源维生素浸种均对小麦发芽有调节作用,都能提高其最终发芽率。但是提高幅度有所差异。用VB6 浸种后的小麦提高幅度最多,VC 次之,VPP 提高幅度最小。同时,4 种外源维生素浸种对小麦种子的出芽速度及芽后长势也有一定的影响。VB6、VC 处理的小麦种子出芽速度最快,萌发后长势最好;VB1 出芽速度相对较慢,VPP 最慢,但都大于对照;VB1 处理长势略高于对照,VPP 处理的小麦长势则低于对照。从整体来看,VB6、VC处理促进效应明显, VB1 次之,而VPP 在某些方面无效甚至产生负效应。此外,相同的维生素处理对不同的品种的种子萌发、生长效果也存在差异,各种维生素作用于川育12 的效应均强于对川育16。进一步对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性进行测定、分析。研究发现:并非所有种类的维生素对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性的提高都有帮助。幼苗根系TTC 还原力在不同维生素处理下存在显著差异,而与小麦品种关系甚微。经VB6、VC 处理后,根系TTC 还原力测定值均显著高于对照,VB1 不明显,VPP 则略低于对照。VB6、VC 处理的幼苗叶片中硝酸还原酶的含量大于对照,VB1 与对照相差无几,而VPP 处理的川育12 幼苗叶片中的硝酸还原酶活性比对照CK 略高,而在川育16 中则略比对照CK 有所下降,呈现出抑制效应。综上结果表明:VB6、VC 具有促进种子发芽,幼苗生长及根系生长的作用,是较好的促生长剂;VPP 具有抑制作用,是较好的抑制剂,可进一步研究、开发利用。在盐胁迫条件下,对用4 种不同维生素VB1、VC、VB6、VPP 浸种川育12、川育16 后的种子萌发及幼苗生长(幼苗的根长、根重、苗高、苗鲜重)的研究结果表明:在不同盐浓度胁迫条件下, 各处理的种子萌发及幼苗生长均受到不同程度的抑制。随着盐浓度的增加, 发芽率、发芽指数和活力指数成下降趋势;幼苗的根长、根重、苗高、苗鲜重不断降低。4 种维生素处理间也表现出较大差异。VB6、VC 在每个处理中均保持对盐害的缓解作用,VB6 较VC 更易于促进发芽及幼苗生长。最终发芽率高,根系多、长、重,苗高高、重。而VB1、VPP 则表现出抑制作用。在高盐浓度150mM 时,4 种维生素浸种后的种子,其最终发芽率均不能达到40%,但VB6、VC 处理最终发芽率、苗重、根重均高于对照,VPP 最终发芽率、苗重、根重均低于对照。进一步对幼苗根系TTC 还原力及幼苗叶片中脯氨酸含量进行测定、分析。研究发现:不同盐浓度,不同维生素处理、不同品种间存在差异。随着盐浓度的增加(75mM,100mM,150mM),幼苗根系TTC 还原力活性成下降趋势,幼苗叶片中脯氨酸的积累量成上升趋势。VB6 处理脯氨酸含量增加最为明显,VC 次之,VPP 与对照接近,其变化幅度最小。经VB6、VC 处理后的幼苗根系还原强度,在不同盐浓度下,测定值均显著高于对照,VB1 不明显,VPP 则低于对照,产生负效应。此外,品种间表现不尽相同,相同的维生素处理,相同的盐浓度对不同的品种的种子萌发、生长效果也存在差异, 4 种维生素对川育16 的作用均强于川育12,但其影响趋势是一致的。说明VB6、VC 具有耐(抗)盐性,可以促进种子发芽和幼苗生长,是较好的耐(抗)盐拌种剂。 Vitamin is one kind of necessary low molecular compound for humans tosustain health and life. Lots of Studies have been done on the effectc of the vitaminsfor people. Vitamin can help people improve the body's natural resistance to disease,Drop the rate of birth defects、cacers and the incidence of the heart diseases. Ifpeople have less of them, the metabolism of the organism may throw off balance,immunity may drop off, and catch disease; Though the effects for Vitamin to thecrops are limited. up to now, there’s no one use soking seeds of wheats with vitaminsas a method, to study on how the effects will happen on the wheat seed germinationand seedling growth, and there are only few reserches on antireversion force forwheats ,none for the antireversion force in Sault stress condition.Wheat(Triticum aestivum L.)is sensitive to the salt, so the salt damage will doharm to wheat’s growth, it will have an unfavorable impact on the output and thequality of wheat.On this reaserch, we Soaking CHY12(red)、CHY16 (white) wheat seeds withVitamin C, B1, PP, B6 (50mg/L) as a pretreatment first. Then under two condition: one is in the normal environment the other is in different Salinity, we begin ourexperiments. Then disscuss on if the vitamin and salinity affect the wheat seedgermination and seedling growth, and what is the different between the two of them,the result shows that:Under the normal condition, after soaking seeds with VB1、VC、VB6、Vpp,we study on the their seed germination and the seeding growth(the root length andweights, The seedling heights and weights), it shows that all of those four kinds ofvitamin can adjust the seed germination, but different in The growth rate. VB6 isbest for increase, VC comes second,VPP is the worst. Meanwhile, those four vitaminalso have effect on the speed of the sprouting of the wheat. VB6、Vc can faster theseed germination most, and the seedlings are all doing well; VB1 do little effects onthe budding, Vpp is the worst, but all treatments are better than CK; but in Vi, VB1some what above the CK, while VPP lower than that. On the whole, the acceleratingeffect of VB6、VC are obvious, VB1 takes second place, but VPP in some aspects arenoneffective even have negative effect. Furthermore, different kind of seeds with thesame vitamin may different in seed germination and seedling growth, four vitaminson CHY16 is better than CHY12.More studies on TTC reductive capacity of roots and the activity of nitratereductase in the leaves, the reasult shows not all the vitamin can help the seedlings toimprove the TTC reductive capacity and the activity of nitrate reductase. TTCreductive capacity in different treatments shows significant differences,but notcorrelate to the variety of the wheat. The TTC reductive capacity of VB6、Vctreatments are all higher than CK, VB1 is nearly the same as CK, VPP is a littlelower than CK. Through the study of acivity of nitrate reductase, it shows that,VB6、VC are higher than CK ,VB1 is nearly the same as CK also, VPP is a little higher inthe CK of CHY12 but lower in CHY16. Through all the results above: VB6、Vc helpthe wheat seed germination, seedling growth and the growth of roots, is theperfectable factor of stimulating the growth; Vpp is a inhibition, that’ll be furtherreserch,and well develop and utilize in the future.Under the different Salinity condition, after soaking seeds with VB1、VC、VB6、Vpp,we study on the their seed germination and the seeding growth(the root lengthand weights, The seedling heights and weights), it shows that: under differentsalinity, the seed germination and the seedling growth of any treatment are inhibited.With the increase of the concentration, the germination rate, Vi、Gi all had fallen; theroot length and weight, the seedling heights and weights steadily sank down. There are also have pronounced difference between all treatments with four differentvitamins.VB6、VC in all treatments are alleviative the salt damage, VB6 is easier tocause to put forth buds than VC, and it’s quantitative value is the highest in theultimate germination rate, in root and seedlings’ hight and weight. Though the VPP、VB1 are seems to inhibite its growth. Under the high concentration150mM Nacl, theultimate germination rate in all treatments are below the 40%, but VB6、VC’squantitative values in any experiments are higher than CK,while VPP lower thanCK.Then we study on the TTC reductive capacity of roots and the content of Polinein leaves, the result shows that between the different salinity, different vitamintreatments, different varieties of the wheat have discrepancy.along with theincreasing concentraion of the salinity(75mM,100mM,150mM),TTC reductivecapacity of roots decreases, the accumulation of the content of Poline in leaves havean upward trend. The increase of VB6’s treatment are obviously, VC comessecond,VPP is nearly come up with CK, changes a little. In TTC reductive capacity of roots’s reserch, VB6、VC are higher than CK at any time,VB1 is not palpable,VPP is lower than CK, makes negative affect on wheat. In addition, varieties of thewheats are remain different, no matter it shows promoting or inhibiting, all fourvitamins have moreobvious effects on CHY16 than CHY12, but the tendency of theeffection are the same. It is say that VB6、VC can help wheat to standwith the saultwell, and promot in growth,they are the better reagent to mix with the seed.
Resumo:
A flow injection system for the determination of organophosphate and carbamate pesticides is described. A sensitive fluorescence probe was synthesized and used as the pH indicator to detect the inhibition of the enzyme acetylcholinesterase (ACNE). The percentage inhibition of enzyme activity is correlated to the pesticide concentration. Several parameters influencing the performance of the system are discussed. The detection limits of 3.5, 50, 12 and 25 mug/l for carbofuran, carbaryl, paraoxon and dichlorvos, in pure water, respectively were achieved with an incubation time of 10 min. A complete cycle of analysis, including incubation time, took 14 min. The detection system has been applied to the determination of carbofuran in spiked vegetable juices (Chinese cabbage and cole), achieving recovery values between 93.2 and 107% for Chinese cabbage juice and 108 and 118% for cole juice at the different concentration levels assayed. (C) 2004 Elsevier B.V. All rights reserved.
Resumo:
PCR扩增了苜蓿根瘤菌乙酰辅酶A合成酶编码基因 (acsA1) ,克隆到连接 -非依赖型载体pET30LIC ;在E .coliBL2 1(DE3)pLysS中得到了有效表达 ,表达需IPTG的诱导 ,诱导 3h达到酶活高峰 .采用His·Bind柱层析对ACS进行了纯化 ,纯化的酶蛋白经SDS-PAGE呈单一浓带 ,分子量约 72 0 0 0 ,具较高的酶活 ,是无细胞提取液的 12 .7倍 .酶动力学分析显示 ,Vmax、Km分别为 (4 13.6± 11.7)mmolL-1和 (5 .8± 0 .6 )mmolL-1.图 4表 2参 8
Resumo:
The mechanism of inhibition of polymer film dewetting is investigated by adding a star comb-like polymer, four-arm P(S-ran-VB-g-PMMA), to PS film and PMMA film on different substrates. It is found that the mechanism of inhibition of polymer film dewetting is kinetic in nature, and is related to the miscibility between the additional compound and the polymer film. On addition to the miscible system [four-arm P(S-ran-VB-g-PMMA) and PMMA], the star comb-like polymers can increase the resistant force of dewetting with hole growth and inhibit the dewetting process of the thin polymer film by enrichment in the rim.
Resumo:
In this paper, a microarray-based surface-enhanced Raman spectroscopic (SERS) assay for detection of kinase functionality and inhibition has been reported. Biotinylated anti-phosphoserinen antibodies mark the phosphorylation and inhibition events and gold nanoparticles are attached to the antibodies by standard avidin-biotin chemistry, followed by silver deposition for SERS signal enhancement. The avidin conjugated fluorescein is used as SERS probe. The alpha-catalytic subunit of cyclic adenosine 5'-monophosphate (cAMP) dependent protein kinase (PKA), its well known substrate, kemptide, and three inhibitors, H89, HA1077, and KN62 have been chosen here to establish the SERS assay. As expected, highly selective inhibition of PKA is demonstrated with the inhibitor H89 and the inhibition assay enable to detect kinase inhibition as well as derive IC50 (half maximal inhibitory concentration) plots.