曙红B及变色酸与核酸作用的光谱学性质研究

利用荧光光谱法、紫外-可见吸收光谱法、圆二色谱法、凝胶电泳法对曙红B及变色酸与核酸的作用机理进行了探讨. 当ct-DNA浓度增加时,曙红B和变色酸的荧光光谱呈荧光增强趋势;紫外-可见光谱呈下降趋势;并使ct-DNA圆二色谱正负峰吸收强度呈现一定的降低;琼脂糖凝胶电泳实验中,二者在泳道中与DNA作用发出的荧光在紫外灯下皆清晰可见. 并将所得结果与溴化乙锭-DNA体系性质进行对比,为寻找高效低毒的DNA分子荧光探针提供了实验基础.     

牛血红蛋白与双十二烷基二甲基溴化铵相互作用的光谱研究(英文)

运用荧光光谱、紫外-可见吸收光谱和圆二色谱法研究了双十二烷基二甲基溴化铵(DDAB)与牛血红蛋白(Hb)的相互作用。从紫外-可见吸收光谱观察到,随着DDAB的浓度增大,Hb在406nm处的特征吸收峰强度下降,且峰位蓝移,说明DDAB导致血红素辅基微观环境变化。由荧光光谱研究可以得出随着DDAB的浓度增大,Hb在340nm处的荧光强度逐渐增强,说明导致色氨酸荧光淬灭的血红素辅基与色氨酸的距离增大。由Scatchard方程计算了不同温度下该反应的表观结合常数、结合位点数及结合热力学参数,热力学参数的变化表明DDAB与Hb之间以疏水作用力为主。圆二色谱的研究进一步表明DDAB使Hb产生轻微的二级结构改变,α-螺旋含量增加.     

酶解小麦蛋白产物-还原糖美拉德反应的光谱研究

采用紫外-可见吸收光谱和荧光光谱研究了酶解小麦蛋白产物与还原糖不同加热条件下的美拉德反应及其产物.美拉德反应在紫外区240和294 nm产生两个特征峰,荧光的最大激发和发射波长为347和450 nm;随反应进行,紫外光吸收和荧光强度迅速增加,表明美拉德反应进入高级阶段,产生的糠醛类、呋喃酮类、吡喃酮类、噻吩类及噻唑类等小分子物质表现较大的积累速率.温度升高,强度增加速率增大.在较高温度时,紫外光吸收出现最大平稳值;荧光强度则到达最大值后降低,表明小分子物质间或与肽聚合生成大分子黑素类物质,小分子物质的积累表现消除速率,反应进入终级阶段.     

含芳香稠环或杂环的光致变色螺吡喃化合物的光谱性能及与DNA作用研究

研究9种带有芳香稠环或杂环基团的光致变色螺吡喃化合物的光致变色性能和荧光性质.通过紫外-可见吸收光谱和荧光光谱对该类化合物与DNA相互作用性能进行了初步探讨.     

碲化镉量子点作为探针研究核黄素与鲑鱼精DNA的相互作用

在水相中合成了巯基丙酸包覆的CdTe量子点（CdTe QDs）,以CdTe QDs作为探针,在pH 7.25 Britton-Robinson（B-R）缓冲溶液中,应用荧光光谱法、紫外吸收光谱法,对核黄素（RF）与鲑鱼精DNA作用方式进行了研究.RF与DNA作用时,使荧光强度降低,紫外吸收明显减色,通过盐效应实验和DN...     

金纳米花的合成及其与人血清白蛋白相互作用的研究

采用晶种法合成了金纳米花（AuNFs）。利用紫外可见吸收光谱法和原子力显微镜方法对金纳米花进行了表征,结果表明制备的金纳米花呈蓝紫色,在549nm处有特征吸收峰,且形状为花状。借助紫外可见吸收光谱法和荧光光谱法研究了AuNFs与人血清白蛋白（HSA）的结合过程。当AuNFs与HSA相互作用时,随着AuNFs溶液浓度的增加,HSA的紫外吸收峰强度也随之增加,但荧光光谱强度则发生了猝灭。由变温荧光实验可以获得两者相互作用的热力学参数,如结合常数（K_a）、吉布斯自由能变（ΔG）、熵变（ΔS）、焓变（ΔH）。ΔG<0表明在溶液中AuNFs与HSA可以自发结合发生反应并形成复合物;ΔH<0、ΔS<0表明结合过程以范德华力和氢键为主。同步荧光和3D荧光实验均表明,AuNFs的存在使得HSA的构象以及周围的环境发生了改变。     

香豆素席夫碱的Zn~(2+)识别研究

本文采用5-氯水杨醛与3-氨基-7-羟基香豆素反应,合成了一种新型的香豆素席夫碱化合物3-[(5-氯-2-羟基-苯亚甲基)-氨基]-7-羟基香豆素(CHB),并采用核磁共振谱、红外光谱和元素分析对合成产物进行了表征。在CHB的DMF溶液中加入Zn~(2+)后,溶液颜色由无色迅速变为橙黄色,紫外-可见吸收光谱最大吸收峰从380nm红移至480nm,而且该分子探针在580nm处的荧光强度显著增强,可通过肉眼观察到其在365nm紫外灯下发出的荧光,而其它离子加入后荧光变化微弱或没有变化,表明该分子探针对Zn~(2+)的选择性较高。吸收光谱的变化表明CHB与Zn~(2+)形成了新的配合物,采用Benesi-Hilderbrand方程计算出两者之间以1∶1配位。     

水溶性卟啉及其系列金属化合物的合成、抗癌活性及其作用机制研究

合成了水溶性四-[(4-三甲铵基)苯基]卟啉TAPP及其金属卟啉化合物MTAPP,其中M=Zn(),Cu(),Fe(),Co(),Ni(),Mn().通过元素分析、紫外-可见光谱确定了化合物的组成和结构.以紫外-可见光谱、荧光光谱及粘度法等手段研究了卟啉化合物TAPP和金属卟啉化合物ZnTAPP与小牛胸腺DNA的相互作用,结果表明,TAPP,ZnTAPP与DNA相互作用使紫外-可见光谱发生了8～12nm的红移,并出现了明显的减色效应.荧光光谱强度明显增大,吸收峰位发生变化.以艾氏腹水癌(Ac)和肝癌(Hept)细胞为靶细胞,采用MTT染色法评价了化合物的抗癌活性,部分化合物表现出较高的抗癌活性     

光谱法研究萘酰亚胺类衍生物与牛血清白蛋白的相互作用

采用荧光光谱、紫外-可见吸收光谱、红外光谱和圆二色光谱研究了3-氨基-4-二甲氨基-N-丁基-1.8-萘酰亚胺(ADBN)与牛血清白蛋白(BSA)结合反应的特征。荧光光谱与紫外-可见吸收光谱表明,ADBN使BSA荧光猝灭的机理为静态猝灭。根据热力学参数判断ADBN与BSA主要是通过氢键和范德华力结合,结合距离为4.08 nm。同步荧光光谱、圆二色光谱以及红外光谱表明,结合过程中BSA的构象发生了变化。     

金属配合物与DNA的弱相互作用

介绍了金属配合物与DNA相互作用的嵌入、沟槽和静电3种主要模式,并介绍了目前常用的研究方法,包括紫外-可见吸收光谱法、荧光光谱、黏度法、循环伏安法。     

Studies of interaction of emodin and DNA in the presence of ethidium bromide by spectroscopic method

Emodin interacting with deoxyribonucleic acid (DNA) has been studied by different spectroscopic techniques, such as fluorescence, ultraviolet and visible (UV-vis), and fourier transform infared (FT-IR) spectroscopies, using ethidium bromide (EB) as a fluorescence probe of DNA. The decrease in the fluorescence of DNA-EB system on addition of emodin shows that the fluorescence quenching of DNA-EB complex by emodin occurs. The binding constants of emodin with DNA in the presence of EB are 6.02x10(4), 9.20x10(4) and 1.17x10(5)Lmol(-1) at 20, 35 and 50 degrees C, respectively. FT-IR spectrum further suggests that both the phosphate groups and the bases of DNA react with emodin. The reaction of DNA with emodin in the presence of EB is affected by ionic strength and temperature. The values of melting temperature (T(m)) of DNA-EB complex and emodin-DNA-EB complexes were determined, respectively. From the experiment evidences, the major binding mode of emodin with DNA should be the groove binding.     

ABSORPTION SPECTRA OF DEOXYRIBOSE, RIBOSEPHOSPHATE, ATP AND DNA BY DIRECT TRANSMISSION MEASUREMENTS IN THE VACUUM-UV (150—190 nm) AND FAR-UV (190—260 nm) REGIONS USING SYNCHROTRON RADIATION AS A LIGHT SOURCE

Transmission measurements of 2-deoxy-D-ribose, D-ribose-5-phosphate, ATP and DNA at 5 nm intervals were made with thin films in the wavelength region between 150 nm and 260 nm using synchrotron radiation. ATP and DNA exhibited two peaks in the absorption spectra around 260 nm and 190 nm, and a steep increase below 170 nm, while ribose phosphate and deoxyribose only exhibited the increase below 190 nm with no appreciable absorption above 190 nm. Since adenine does not exhibit the increase of absorption below 180 nm, these results indicate that the absorption of the sugar-phosphate group, rather than adenine, contributed to the increase below 170 nm in the absorption spectra of ATP and DNA.     

FLUORESCENT TAUTOMERS AND THE APPARENT PHOTOPHYSICS OF ADENINE AND GUANINE

Abstract— Fluorescence, absorption and fluorescence excitation spectra, and quantum yields of 0.02 mM solutions of adenine, 7-methyladenine (7-MA), guanine and 7-melhylguanine (7-MG) are presented for excitation with240–300 nm light. The solvent is neutral ethylene glycol-water (70:30 v/v) in the temperature range140–165 K. Phosphorescence spectra of adenine and 7-MA at 140 K are also presented. The excitation spectrum of adenine shows vibrational structure, whereas the absorption does not. However, the fluorescence of adenine shows the vibrational structure, as do the absorption, fluorescence and excitation spectra of 7-MA. The results confirm (and reinforce) the notion that luminescence from adenine under these conditions is from the N₇–H tautomer, instead of the more abundant N₉–H form. In a similar fashion, the data from guanine and 7-MG strongly suggest that the luminescence from guanine is also mostly from the N₇–H tautomer.     

In vivo monitoring the changes of interstitial pH and FAD/NADH ratio by fluorescence spectroscopy in healing skin wounds

The aim of our study was to evaluate the changes of interstitial pH and flavin adenine dinucleotide (FAD)/reduced nicotinamide adenine dinucleotide (NADH) ratio in healing skin wounds using fluorescence spectroscopy in Sprague Dawley rats. In the experiment, excisional and incisional models of wound healing were used. The florescein as the pH-sensitive probe using excitation spectra (lambda(Em) = 535 nm) was used for the measurement of pH changes, and synchronous fluorescence spectra (Deltalambda = 60 nm) for the monitoring of FAD/NADH ratio changes were measured from the surfaces of healing wounds. Increase of interstitial pH and FAD/NADH ratio was recorded during the time interval from the 15th to the 65th minute after surgery. The decrease of pH between the 48th and the 72nd hour after surgery as well as the increase of FAD/NADH ratio between the 72nd and the 96th hour of wound healing were recorded. The results indicate that the use of fluorescence spectroscopy may be considered as a valuable tool for noninvasive in vivo monitoring of selected redox parameters in the early phases of wound healing.     

Quadracyclic Adenine: A Non‐Perturbing Fluorescent Adenine Analogue

Fluorescent‐base analogues (FBAs) comprise a group of increasingly important molecules for the investigation of nucleic acid structure and dynamics as well as of interactions between nucleic acids and other molecules. Here, we report on the synthesis, detailed spectroscopic characterisation and base‐pairing properties of a new environment‐sensitive fluorescent adenine analogue, quadracyclic adenine (qA). After developing an efficient route of synthesis for the phosphoramidite of qA it was incorporated into DNA in high yield by using standard solid‐phase synthesis procedures. In DNA qA serves as an adenine analogue that preserves the B‐form and, in contrast to most currently available FBAs, maintains or even increases the stability of the duplex. We demonstrate that, unlike fluorescent adenine analogues, such as the most commonly used one, 2‐aminopurine, and the recently developed triazole adenine, qA shows highly specific base‐pairing with thymine. Moreover, qA has an absorption band outside the absorption of the natural nucleobases (>300 nm) and can thus be selectively excited. Upon excitation the qA monomer displays a fluorescence quantum yield of 6.8 % with an emission maximum at 456 nm. More importantly, upon incorporation into DNA the fluorescence of qA is significantly less quenched than most FBAs. This results in quantum yields that in some sequences reach values that are up to fourfold higher than maximum values reported for 2‐aminopurine. To facilitate future utilisation of qA in biochemical and biophysical studies we investigated its fluorescence properties in greater detail and resolved its absorption band outside the DNA absorption region into distinct transition dipole moments. In conclusion, the unique combination of properties of qA make it a promising alternative to current fluorescent adenine analogues for future detailed studies of nucleic acid‐containing systems.     

Spectroscopy studies of sandwich-type complex of silver(I) co-ordinated to nuclear fast red and adenine and its analytical applications

A novel fluorometric method for the determination of adenine was proposed, based on the fluorescence quenching of a nuclear fast red-silver complex as a small molecule optical probe in the presence of adenine. The characteristic spectra of absorption, resonance Rayleigh scattering (RRS) and the fluorescence of the system were analysed. Adenine, Ag⁺ and nuclear fast red formed a sandwich-type complex. The composition of the compound and the mechanisms of fluorescence quenching were discussed. The optimal conditions for the reaction and the effects of co-existing substances were also investigated. The quenched fluorescence intensity was proportional to the concentration of adenine in the range of 0.03–0.90 µg mL^?1, with the limit of detection of 0.023 µg mL^?1. The method was applied satisfactorily to the determination of adenine in synthetic samples, herring sperm DNA, human sera and vitamin B₄ tablets with recoveries from 95.3 % to 105 %.     

Quadracyclic adenine: a non-perturbing fluorescent adenine analogue

Fluorescent-base analogues (FBAs) comprise a group of increasingly important molecules for the investigation of nucleic acid structure and dynamics as well as of interactions between nucleic acids and other molecules. Here, we report on the synthesis, detailed spectroscopic characterisation and base-pairing properties of a new environment-sensitive fluorescent adenine analogue, quadracyclic adenine (qA). After developing an efficient route of synthesis for the phosphoramidite of qA it was incorporated into DNA in high yield by using standard solid-phase synthesis procedures. In DNA qA serves as an adenine analogue that preserves the B-form and, in contrast to most currently available FBAs, maintains or even increases the stability of the duplex. We demonstrate that, unlike fluorescent adenine analogues, such as the most commonly used one, 2-aminopurine, and the recently developed triazole adenine, qA shows highly specific base-pairing with thymine. Moreover, qA has an absorption band outside the absorption of the natural nucleobases (>300?nm) and can thus be selectively excited. Upon excitation the qA monomer displays a fluorescence quantum yield of 6.8?% with an emission maximum at 456?nm. More importantly, upon incorporation into DNA the fluorescence of qA is significantly less quenched than most FBAs. This results in quantum yields that in some sequences reach values that are up to fourfold higher than maximum values reported for 2-aminopurine. To facilitate future utilisation of qA in biochemical and biophysical studies we investigated its fluorescence properties in greater detail and resolved its absorption band outside the DNA absorption region into distinct transition dipole moments. In conclusion, the unique combination of properties of qA make it a promising alternative to current fluorescent adenine analogues for future detailed studies of nucleic acid-containing systems.     

Photochemistry and phototoxicity of aloe emodin

Photochemical pathways leading to the phototoxicity of the aloe vera constituent aloe emodin were studied. The results indicate a photochemical mechanism involving singlet oxygen to be the most likely pathway responsible for the observed phototoxicity. Aloe emodin was found to efficiently generate singlet oxygen when irradiated with UV light (phidelta = 0.56 in acetonitrile). The survival of human skin fibroblast cells in the presence of aloe emodin was found to decrease upon irradiation with UV light. A further decrease in cell survival was observed in D2O compared with H2O, suggesting the involvement of singlet oxygen as the primary pathway. Laser flash photolysis experiments were also carried out on aloe emodin alone and in the presence of various biological substrates. Aloe emodin proved to be relatively photostable (phi = 1 x 10(-4)) and a poor photo-oxidant (E*red = +1.02 V). Only absorption bands caused by the triplet state of aloe emodin (lambdamax = 480 nm) and the aloe emodin conjugate base (lambdamax = 520 nm) were observed in the transient spectra.     

Microanatomical and biochemical origins of normal and precancerous cervical autofluorescence using laser-scanning fluorescence confocal microscopy

Clinical studies have shown that in vivo fluorescence spectroscopy can improve the diagnosis of cervical precancer. Recent work suggests that epithelial fluorescence increases, whereas stromal fluorescence decreases, with precancer. However, the microanatomic and biochemical sources of fluorescence in living cervical tissue have not yet been established. This study aims to characterize the origins of living normal and precancerous cervical fluorescence at microscopic levels using laser-scanning fluorescence confocal microscopy. Ten pairs of colposcopically normal and abnormal biopsies were obtained; transverse, 200 microm thick, short-term tissue cultures were prepared and imaged when viable with UV (351-364 nm) and 488 nm excitation before and after addition of the vital dye, Mitotracker Orange. In normal epithelium basal epithelial cells showed cytoplasmic fluorescence; parabasal, intermediate and superficial cells showed fluorescence only at the periphery of the cell. In low-grade precancers cytoplasmic fluorescence was visible in the bottom one-third of the epithelium; in high-grade precancers cytoplasmic fluorescence was visible throughout the lower two-thirds of the epithelium. Cytoplasmic fluorescence was colocalized with the MitoTracker probe and is attributed to mitochondrial reduced form of nicotinamide adenine dinucleotide at UV excitation and mitochondrial flavin adenine dinucleotide at 488 nm excitation. Stromal fluorescence originated from matrix fibers; with the development of precancer the density and fluorescence intensity of matrix fibers decrease. Autofluorescence properties of precancerous cervix reflect an increased number of metabolically active mitochondria in epithelial cells and a reduced stromal fluorescence, which can be an indicator for altered communication between precancerous epithelium and stroma. These changes can explain differences in in vivo fluorescence spectra of normal and precancerous cervical tissue.     

S-异丙甲草胺与小牛胸腺DNA的相互作用

应用紫外光谱、荧光光谱、DNA热变性法以及黏度法研究了S-异丙甲草胺与小牛胸腺DNA(ctDNA)的相互作用. 结果表明, S-异丙甲草胺使ctDNA在200 nm处的吸收峰发生明显改变, 表现出红移和减色效应, 而对260 nm处的吸收峰产生影响较小, 排除了嵌插作用的可能; ctDNA对S-异丙甲草胺内源性荧光表现出很强的猝灭作用, 且随温度的升高, 其猝灭程度有所下降, 表明S-异丙甲草胺是以形成加合物的方式与ctDNA结合的, 并求得了它们在不同温度下的结合常数; 将不同离子强度条件下S-异丙甲草胺与ctDNA作用以及不同S-异丙甲草胺浓度下ctDNA的热变性温度和黏度变化的研究结果与紫外光谱和荧光光谱相结合, 可以判断S-异丙甲草胺是以沟槽作用的方式与ctDNA结合的.