Triton X-100在单菌落PCR技术中的优化作用

在利用单菌落PCR法直接筛选含有外源基因的重组质粒以及发生同源重组的重组子时,通过0.1%TritonX-100处理模板对PCR技术进行优化。以大肠杆菌DH5α、单核细胞增生李斯特菌的重组克隆为例,单菌落经Triton X-100处理后再进行PCR,琼脂糖凝胶电泳图谱的条带更清晰、杂带减少;克隆鉴定的阳性率明显提高,且在15μL的PCR反应混合液体系中的扩增效果更明显。对单核细胞增生李斯特菌的检测方法可以推广适用于多种革兰氏阳性菌。     

Triton X-100反相微乳液体系制备纳米粒子的研究进展

介绍了Triton X-100反相微乳液体系的形成机理、特性和制备纳米粒子的原理,总结了采用Triton X-100体系制备纳米粒子的影响因素。综述了近年来国内外用该微乳液系统制备纳米粒子的新进展,提出了该体系制备纳米颗粒存在的问题。     

在TritonX－100存在下用2－QADN2，7示波极谱法测定痕量铜

合成了１－（２－喹啉偶氮）－２，７－二羟萘，研究了本试剂与铜络合物的极谱特性。在ｐＨ９．６０的缓冲溶液中，当有０．００２５％ＴｒｉｔｏｎＸ－１００存在时，Ｃｕ（Ⅱ）－２－ＱＡＤＮ２，７络合物在－０．４１Ｖ产生一灵敏示波极谱波，用此波检测痕量铜的下限为１．０×１０－１０ｍｏｌ／Ｌ。     

Measurement of Hydrogen Produced during Magnesium Corrosion in Hydrochloric Acid and the Effect of the Triton X-100 Surfactant on Hydrogen Production

The goal of this article is to measure hydrogen produced during the corrosion of magnesium in HCl and the influence of the Triton X-100 surfactant on hydrogen production. It was found that the hydrogen produced during corrosion of Mg in HCl increased with increasing HCl concentration, stirring rate, temperature, and time of immersion. The addition of the Triton X-100 surfactant inhibits the amount of hydrogen evolved. The inhibition behavior was explained on the basis of adsorption of Triton X-100 molecules on the Mg surface creating a barrier for mass and charge transport, which protects the Mg surface from aggressive ions. The activation thermodynamic parameter values were calculated and explicated. Some theoretical chemical parameters were also calculated. The results obtained from the theoretical calculations are in agreement with the practical results.     

Apparent electrocatalysis on 3D nanoporous platinum film electroplated from hexagonal lyotropic liquid crystalline phase of Triton X-100

Electrocatalytic effect of nanoporous interface between platinum (Pt) and electrolyte was investigated in terms of direct electrochemical oxidation of glucose. A Pt film with 3D nanopores was electroplated from the hexagonal (H₁) lyotropic liquid crystal (LLC) phase of t-octylphenoxypolyethoxyethanol (Triton X-100), a nonionic surfactant with a phenyl group in a hydrophobic tail. This Pt film, which was electroplated from H₁-LLC of Triton X-100 (ePt-H₁-TX100), comprised closely stacked crystalline (face centered cubic) nanoparticles (diameter, 4-7 nm) with 3D nanopores (width, 1-2 nm) developed among the particles. The ePt-H₁-TX100 showed high surface roughness and selective enhancement of a sluggish redox reaction, that is, kinetic-controlled reaction such as glucose oxidation. Nonenzymatic glucose sensing can be achieved by using discriminatively enhanced sensitivity to glucose over redox active interfering molecules.     

Effect of Triton X-100 in water-added electrolytes on the performance of dye-sensitized solar cells

The performance of a Ru(II) dye-sensitized nanocrystalline TiO₂ solar cell (DSSC) with both Triton X-100 and water as additives in 3-methoxypropionitrile solutions is described. In the presence of these additives, the open-circuit voltage (V_oc), fill factor and stability of the cell increased, while the short-circuit photocurrent density decreased compared to those found in their absence. Based on these changes, the DSSC yielded a higher solar-to-electricity conversion efficiency of 5.9%, compared to 5.3% for the cells fabricated without the additives. Furthermore, the addition of Triton X-100 to the water-added electrolyte was found to render the Ag metal corrosion resistant.     

Interaction of triton X-100 on silica: A relationship between surface characteristics and adsorption isotherms

Adsorption of Triton X-100 on various silica substrates has been investigated. A number of solids, including a natural quartz, this quartz washed with HCl acid and subsequently heated at 1273 K; two aerosils and one Kieselgel silicas were studied. These solids exhibit surface areas in the range of 5 to 430 m² g^?1. All the Triton adsorption isotherms display an S-shape at the adsorption temperatures studied (298 and 308 K). It has been found that the pretreatments of natural quartz (by water washing, impurities removed by acid and/or high temperature calcination) affect considerably the amounts of TX-100 adsorbed. Measurements of surface composition have been made by X-ray photoelectron spectroscopy (XPS) with particular emphasis on the presence of impurities and on the number of OH groups at the surface of the samples. The nature of the surface hydroxyl has also been studied by infrared spectroscopy. Furthermore, the specific number of hydroxyl groups on the surface of the silica samples has been determined by thermogravimetric analysis. Finally an attempt to correlate solid surface characteristics with adsorption isotherms has been developed.     

曲拉通增敏的分光光度法测定水中挥发酚类

在pH=10.0±0.2的氨缓冲介质中,苯酚被(NH4)2S2O8氧化生成醌类,与4-氨基安替比林(4-AAP)反应生成橙红色的吲噪苯酚安替比林染料.加入非离子表面活性剂Triton X-100,增敏增稳,λmax=490 nm,体系在2 h内获得稳定光度值,3 h内误差小于±5%.最低检出限浓度为0.1 mg·L-1.苯酚浓度在0.5～14.5μg·mL-1范围内符合比尔定律,其线性方程为A=0.0053 12416c,ε=1.25×104L·mol-1·cm-1,RSD=3.97‰.用于挥发酚类污水样的测定,结果令人满意.     

Fate of Triton X-100 Applications on Water and Soil Environments: A Review

Triton X-100 applications as surfactant raises concern on water and soil environment due to its non-biodegradability and inhibition effect. This paper aims at reviewing Triton X-100 biodegradability and inhibition literature. It shows Triton X-100 is biodegradable by aerobic and anaerobic municipal wastewater sludge and Vibrio cyclitrophicus-sp-Nov organism. Adsorption and biodegradation are mechanisms of Triton removal. Triton inhibits anaerobic sludge organisms and some single aerobic organisms. Inhibition mechanisms are substrate shortage, physiological membrane-damaging and/or alteration in organism cell membrane. Thus Triton X-100 fate in the environment and its sustainable application can be controlled via proper selection of organism type, Triton concentration, and substrate.     

Supported Lipid Bilayer Platform for Characterizing the Membrane-Disruptive Behaviors of Triton X-100 and Potential Detergent Replacements

Triton X-100 (TX-100) is a widely used detergent to prevent viral contamination of manufactured biologicals and biopharmaceuticals, and acts by disrupting membrane-enveloped virus particles. However, environmental concerns about ecotoxic byproducts are leading to TX-100 phase out and there is an outstanding need to identify functionally equivalent detergents that can potentially replace TX-100. To date, a few detergent candidates have been identified based on viral inactivation studies, while direct mechanistic comparison of TX-100 and potential replacements from a biophysical interaction perspective is warranted. Herein, we employed a supported lipid bilayer (SLB) platform to comparatively evaluate the membrane-disruptive properties of TX-100 and a potential replacement, Simulsol SL 11W (SL-11W), and identified key mechanistic differences in terms of how the two detergents interact with phospholipid membranes. Quartz crystal microbalance-dissipation (QCM-D) measurements revealed that TX-100 was more potent and induced rapid, irreversible, and complete membrane solubilization, whereas SL-11W caused more gradual, reversible membrane budding and did not induce extensive membrane solubilization. The results further demonstrated that TX-100 and SL-11W both exhibit concentration-dependent interaction behaviors and were only active at or above their respective critical micelle concentration (CMC) values. Collectively, our findings demonstrate that TX-100 and SL-11W have distinct membrane-disruptive effects in terms of potency, mechanism of action, and interaction kinetics, and the SLB platform approach can support the development of biophysical assays to efficiently test potential TX-100 replacements.     

竹红菌乙素在Triton X-100胶束中的光谱性质及增溶效应研究

为了测定竹红菌乙素(HB)在Triton X-100(TX-100)胶束中的光谱性质及增溶情况,采用紫外-可见分光光度计和荧光分光光度计得到HB在该体系下的吸收光谱和荧光发射光谱,通过HB荧光强度的变化得到增溶情况。在TX-100胶束体系中,竹红菌乙素吸收峰的位置稍有红移且峰型变宽,且荧光发射峰的峰型和峰位均没有大的变化,但最大发射峰强度明显降低。在TX-100胶束的临界胶束浓度(CMC)为58.5μmol·L-1时,HB的最大增溶浓度为50μmol·L-1,得到HB在低浓度TX-100胶束体系中的增溶曲线。该测定方法能够为简单生物模型体系提供一些基础参考数据。     

Triton X-114萃取法去除脑膜炎球菌荚膜多糖中的内毒素

目的采用Triton X-114萃取法去除A群脑膜炎球菌荚膜多糖中的内毒素。方法1% Triton X-114与多糖溶液在0℃混合均匀,分别加热至25、37和56℃,观察分层情况。离心后收集上层水相,对其中残余萃取剂Triton X-114进行去除方法的选择。比较多糖浓度对萃取的影响及二次萃取的效果,并对终产物进行全面检定。对W135群和Y群脑膜炎球菌多糖进行萃取效果比较。结果最终选择56℃10min作为升温条件,3000r/min离心10min后,上清中多糖的内毒率含量均减少85%以上,多糖回收率不低于80%。残余的Triton X-114选择透析法去除。多糖浓度越低,越易于萃取。二次萃取多糖回收率大于85%。内毒素含量可降低至0.706EU/μg多糖。经检测,终产物多糖的相对分子质量未发生改变,免疫原性与萃取前差异无显著意义,异常毒性、多糖及内毒素含量合格。蛋白含量稍有增加,核酸含量降低。W135群和Y群脑膜炎球菌多糖的萃取结果与A群多糖相似。结论Triton X-114萃取法可以用于去除脑膜炎球菌多糖中的内毒素。     

Kinetics and mechanism of adsorption of triton x-100 on a reservoir sand

Adsorption of Triton X-100 on a reservoir sand (characterised by chemical analysis and i.r. spectra to be silica) has been studied under field conditions. The adsorption behaviour is best characterised by Freundlich and Temkin adsorption models. The kinetics of adsorption is found to be very much dependent on temperature and the rate of adsorption shows an anti-Arrhenius temperature dependence. The adsorption is proposed to proceed through Si—CH₃ bond formation. Addition of common inorganic salts is found to cause maxima and minima in the adsorption isotherm and to change the shape of the isotherm.     

Cloud Point Extraction of Four Triphenylmethane Dyes by Triton X-114 as Nonionic Surfactant

A method for removing four triphenylmethane dyes from wastewater by cloud point extraction with the nonionic surfactant Triton X-114 (TX-114) was developed. The triphenylmethane dyes were crystal violet, ethyl violet, malachite green and brilliant green. The cloud point of TX-114 generally increased in the presence of any of the four dyes. In the cloud point system, these dyes were solubilized into a coacervate phase that left a color-free dilute phase. The extraction efficiency of the dyes increased with the temperature, TX-114 concentration, and salt (NaCl and CaCl₂) concentration. More than 97% TX-114 in the dilute phase was recovered by adjusting the volume ratio of dichloromethane to the dilute phase. The Langmuir-type adsorption isotherm was used to describe the dye solubilization. The Langmuir constants m and n were calculated as functions of temperature. The results showed that the solubilization of the triphenylmethane dyes in the cloud point system was related to the partition coefficient and their molecular structures.     

羟基肉桂酸衍生物的合成及其与人血清白蛋白的相互作用

以咖啡酸、间羟基肉桂酸为母体,分别与止血芳酸反应,设计合成两种羟基肉桂酸衍生物I、II,并用MS、IR、1H-NMR、13C-NMR进行结构表征。采用荧光光谱法、同步荧光光谱法、紫外可见光谱法结合分子对接技术研究其在体外生理条件下与HSA之间的相互作用机制。光谱实验结果表明,衍生物I和II都可以使HSA的内源荧光发生静态猝灭,相互作用力主要是氢键和范德华力,且对HSA的构象产生一定的影响。分子对接结果表明衍生物I、II与HSA中的结合位于亚结构域IIA（即site I）中,作用力主要是氢键和范德华力,同时还存在着疏水作用,且衍生物距色氨酸残基(Trp214)很近,很好地解释了衍生物有效猝灭HSA内源荧光的实验现象。分子对接与实验获得了一致性的结果,二者相互作证,提高了实验结果推断的准确度。     

羟基肉桂酸衍生物的合成及其与人血清白蛋白的相互作用

以咖啡酸、间羟基肉桂酸为母体,分别与止血芳酸反应,设计合成两种羟基肉桂酸衍生物(E)-4-{[1'-(3″,4″-二羟基苯基)丙烯酰氨基]甲基}苯甲酸(Ⅰ)、(E)-4-{[1'-(3″-羟基苯基)丙烯酰氨基]甲基}苯甲酸(Ⅱ),并用MS、IR、1HNMR、13CNMR进行结构表征。采用荧光光谱法、同步荧光光谱法、紫外可见光谱法结合分子对接技术研究其在体外生理条件下与人血清白蛋白(HSA)之间的相互作用机制。光谱实验结果表明,衍生物Ⅰ和Ⅱ都可以使HSA的内源荧光发生静态猝灭,相互作用力主要是氢键和范德华力,且对HSA的构象产生一定影响。分子对接结果表明,衍生物Ⅰ、Ⅱ与HSA结合在亚结构域ⅡA(即site I)中,作用力主要是氢键和范德华力,同时还存在着疏水作用,且衍生物距色氨酸残基(Trp214)很近,很好地解释了衍生物有效猝灭HSA内源荧光的实验现象。分子对接与实验获得了一致性的结果,二者相互佐证,提高了实验结果推断的准确度。     

Cholesterol biosensor based on electrochemically prepared polyaniline conducting polymer film in presence of a nonionic surfactant

Cholesterol biosensor has been fabricated by covalently coupling cholesterol oxidase (ChOx) via glutaraldehyde onto electrochemically prepared polyaniline film in presence of TritonX-100 [4-(1,1,3,3-tetramethylbutyl) phenyl polyethylene glycol], a non-ionic surfactant onto indium-tin-oxide (ITO) glass substrate. These ChOx/PANI-TX-100/ITO bioelectrodes have been characterized using Fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry (CV) and scanning electron microscopy (SEM) techniques. The results of response measurements carried out on ChOx/PANI-TX-100/ITO bioelectrodes using amperometric and photometric techniques, reveal detection limit as 5 mg/dl, linearity from 5 to 400 mg/dl of cholesterol and sensitivity as 131 μA/(mg/dl cm⁻²). These biosensing electrodes are thermally stable up to 65 °C, can be used about 20 times and have a shelf-life of about 10 weeks when stored at 4 °C. Attempts have also been made to utilize the ChOx/PANI-TX-100/ITO bioelectrodes for estimation of free cholesterol concentration in serum samples.     

光谱法研究壳聚糖铜与人血清白蛋白的相互作用

通过热回流制备了壳聚糖-铜(CTS-Cu)配合物,采用荧光、红外、紫外光谱对配合物CTS-Cu与HSA的相互作用进行了表征,结果表明:在280nm处出现紫外吸收峰,在340nm处使HSA的内源荧光发生猝灭,猝灭速率常数kq=3.014×1014L/mol·s,猝灭类型为静态猝灭,说明CTS-Cu与HSA发生了较强烈的相互作用,形成了基态复合物,其结合常数Ka=4.27×106L/mol。通过加入位点标记物布洛芬(Ibuprofen)和华法林(Warfarin)用荧光法研究了CTS-Cu与HSA的结合位点,表明Warfarin与CTSCu竞争HSA的同一个位点,即SiteⅠ位。采用微量热法研究了CTS-Cu与核桃内生菌的相互作用,随着CTS-Cu浓度的增加,最大产热效率出现的时间逐渐延长,表明CTS-Cu对核桃内生菌的生长有一定的抑制作用。     

Tumor Cell Capture Using Platelet-Based and Platelet-Mimicking Modified Human Serum Albumin Submicron Particles

The co-localization of platelets and tumor cells in hematogenous metastases has long been recognized. Interactions between platelets and circulating tumor cells (CTCs) contribute to tumor cell survival and migration via the vasculature into other tissues. Taking advantage of the interactions between platelets and tumor cells, two schemes, direct and indirect, were proposed to target the modified human serum albumin submicron particles (HSA-MPs) towards tumor cells. HSA-MPs were constructed by the Co-precipitation–Crosslinking–Dissolution (CCD) method. The anti-CD41 antibody or CD62P protein was linked to the HSA-MPs separately via 1-ethyl-3-(-3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) EDC/NHS chemistry. The size of modified HSA-MPs was measured at approximately 1 µm, and the zeta potential was around −24 mV. Anti-CD41-HSA-MPs adhered to platelets as shown by flowcytometry and confocal laser scanning microscopy. In vitro, we confirmed the adhesion of platelets to tumor lung carcinoma cells A549 under shearing conditions. Higher cellular uptake of anti-CD41-HSA-MPs in A549 cells was found in the presence of activated platelets, suggesting that activated platelets can mediate the uptake of these particles. RNA-seq data in the Cancer Cell Lineage Encyclopedia (CCLE) and The Cancer Genome Atlas (TCGA) database showed the expression of CD62P ligands in different types of cancers. Compared to the non-targeted system, CD62P-HSA-MPs were found to have higher cellular uptake in A549 cells. Our results suggest that the platelet-based and platelet-mimicking modified HSA-MPs could be promising options for tracking metastatic cancer.     

Spontaneous and Ionizing Radiation-Induced Aggregation of Human Serum Albumin: Dityrosine as a Fluorescent Probe

The use of spectroscopic techniques has shown that human serum albumin (HSA) undergoes reversible self-aggregation through protein–protein interactions. It ensures the subsequent overlapping of electron clouds along with the stiffening of the conformation of the interpenetrating network of amino acids of adjacent HSA molecules. The HSA oxidation process related to the transfer of one electron was investigated by pulse radiolysis and photochemical methods. It has been shown that the irradiation of HSA solutions under oxidative stress conditions results in the formation of stable protein aggregates. The HSA aggregates induced by ionizing radiation are characterized by specific fluorescence compared to the emission of non-irradiated solutions. We assume that HSA dimers are mainly responsible for the new emission. Dityrosine produced by the intermolecular recombination of protein tyrosine radicals as a result of radiolysis of an aqueous solution of the protein is the main cause of HSA aggregation by cross-linking. Analysis of the oxidation process of HSA confirmed that the reaction of mild oxidants (Br2•−, N3•, SO4•−) with albumin leads to the formation of covalent bonds between tyrosine residues. In the case of ^•OH radicals and partly, Cl2•−, species other than DT are formed. The light emission of this species is similar to the emission of self-associated HSA.