十二羰基三铁与DNA相互作用的紫外和荧光光谱研究

采用紫外-可见吸收光谱、荧光光谱等法研究了十二羰基三铁簇合物与小牛胸腺DNA的相互作用.在簇合物存在下,DNA的紫外吸收光谱产生了明显的增色效应.荧光光谱表明簇合物的荧光强度随DNA的加入其荧光强度增加,说明簇合物与DNA之间发生了插入作用.     

氢氯噻嗪与DNA相互作用的光谱研究

用紫外光谱法和荧光光谱法研究了氢氯噻嗪(HCT)和小牛胸腺DNA(ctDNA)之间的相互作用.在pH 7.0的磷酸盐缓冲溶液中,HCT的荧光激发峰和发射峰分别位于278nm和360nm处.ctDNA的加入对HCT的荧光存在着很强的荧光猝灭作用,这种荧光静态猝灭作用是由ctDNA和HCT键合引起,键合常数为1.12×10-4L/mol(25℃).采用紫外光谱,离子强度的影响和Ⅰ-猝灭等条件实验研究了HCT与DNA间的相互作用.DNA浓度的变化不改变它们的作用,属于沟槽作用模式.     

紫外辐射对小牛胸腺DNA水溶液影响的拉曼光谱研究

报道了小牛胸腺DNA水溶液经9,20,40min紫外辐射的拉曼光谱图,紫外线的辐射照度为1868W·m-2。实验结果表明波长为2537nm的紫外光在起主要作用的紫外辐射对DNA的损伤是严重的,短短10min的紫外辐射就使1094cm-1这个强峰分裂成几个小峰,说明DNA的构象受到破坏,DNA的构型发生变化,部分单双键发生了断裂,出现了各种各样由于DNA键断裂产生的多核苷酸;4种碱基均受到不同程度的影响,碱基间的氢键造成断裂,其中嘧啶、嘌呤碱基受到的损伤较为严重;紫外辐射对脱氧核糖也产生了破坏。另外,该实验也表明,在水溶液中,DNA以B型结构为主,局部的A型结构仍然存在。     

Interactions of Nile Blue with Micelles, Reverse Micelles and a Genomic DNA

In this contribution we report studies on the nature of binding of a small ligand/drug Nile blue (NB) with sodium dodecyl sulfate (SDS) micelles, bis-(2-ethylehexyl) sulfosuccinate (AOT)/isooctane reverse micelles (RM) and a genomic DNA extracted from Salmon sperm. With detailed steady state and picosecond resolved optical spectroscopic techniques, we examined the fluorescence quenching of the ligand upon complexation with the SDS monomers and DNA. Polarization analyzed picosecond-resolved fluorescence measurements reveal geometrical restriction on the probe in SDS micelles, AOT-RM and DNA. Steady state and time resolved studies on the probe in nanocages of AOT RM with various degrees of hydration (w₀) reveal the existence of NB as two distinct species namely, neutral and cationic. This study confirms that the emission of NB in aqueous micelles and DNA solution is due to the cationic form of the drug. Our experiments clearly identified non-specific electrostatic and intercalative modes of interaction of the probe with the DNA at lower and higher DNA concentrations respectively. The nature of binding of NB in presence of the DNA and SDS micelles reveals that the binding affinity of the probe is higher with the micelles than with the DNA. The complex rigidity of NB with DNA and its fluorescence quenching with DNA elucidate a strong recognition mechanism between NB and DNA.     

Electron microscopy studies on DNA recognition by DNA-PK

Advances in transmission electron microscopy coupled to increasingly powerful biocomputing techniques are opening enormous possibilities to understand the structure and function of complex biological processes performed by large multi-protein assemblies. This is an exciting time for electron microscopists because we can combine our efforts with X-ray crystallographers and NMR spectroscopists to reach the prospect of studying the structure and dynamics of the so-called ‘molecular machines’. One of these fascinating systems is the macromolecular complex formed around double-stranded DNA breaks (DSBs). Non-homologous end-joining (NHEJ) is the main DSBs repair pathway in mammalian cells, where a collection of proteins interact to rejoin two broken DNA ends. During NHEJ, DNA-dependent protein kinase (DNA-PK) binds damaged DNA with high affinity and acts as the main scaffold for other repair factors. Several studies have made use of the electron microscope to reveal the three-dimensional architecture of DNA-PK and the structural basis for the recognition of damaged DNA and the activation of DNA-PK's kinase activity.     

Biomolecular Release Stimulated by Electrochemical Signals at a Very Small Potential Applied

DNA release electrochemically stimulated by applying ?10 mV on the modified electrode was studied. The release process was based on the local (interfacial) pH change produced upon H₂O₂ reduction electrocatalyzed by the immobilized microperoxidase‐11. SiO₂ nanoparticles attached to the electrode surface and functionalized with trigonelline and boronic acid species changed their electrical charge from positive to negative upon the interfacial pH change, thus allowing electrostatic adsorption of negatively charged DNA on the positive interface and then its repulsion/release from the negative interface. The loaded/released DNA molecules were labeled with a fluorescent dye to allow easy detection of the released DNA molecules. The important feature of the developed system is the controlled DNA release upon applying very small electrical potential on the modified electrode.     

Melatonin Rescues the Dendrite Collapse Induced by the Pro-Oxidant Toxin Okadaic Acid in Organotypic Cultures of Rat Hilar Hippocampus

The pro-oxidant compound okadaic acid (OKA) mimics alterations found in Alzheimer’s disease (AD) as oxidative stress and tau hyperphosphorylation, leading to neurodegeneration and cognitive decline. Although loss of dendrite complexity occurs in AD, the study of this post-synaptic domain in chemical-induced models remains unexplored. Moreover, there is a growing expectation for therapeutic adjuvants to counteract these brain dysfunctions. Melatonin, a free-radical scavenger, inhibits tau hyperphosphorylation, modulates phosphatases, and strengthens dendritic arbors. Thus, we determined if OKA alters the dendritic arbors of hilar hippocampal neurons and whether melatonin prevents, counteracts, or reverses these damages. Rat organotypic cultures were incubated with vehicle, OKA, melatonin, and combined treatments with melatonin either before, simultaneously, or after OKA. DNA breaks were assessed by TUNEL assay and nuclei were counterstained with DAPI. Additionally, MAP2 was immunostained to assess the dendritic arbor properties by the Sholl method. In hippocampal hilus, OKA increased DNA fragmentation and reduced the number of MAP2(+) cells, whereas melatonin protected against oxidation and apoptosis. Additionally, OKA decreased the dendritic arbor complexity and melatonin not only counteracted, but also prevented and reversed the dendritic arbor retraction, highlighting its role in post-synaptic domain integrity preservation against neurodegenerative events in hippocampal neurons.     

O6-Alkylguanine DNA Alkyltransferase Mediated Disassembly of a DNA Tetrahedron

Tetrahedron DNA structures were formed by the assembly of three-way junction ( TWJ ) oligonucleotides containing O⁶-2′-deoxyguanosine-alkylene-O⁶-2′-deoxyguanosine (butylene and heptylene linked) intrastrand cross-links (IaCLs) lacking a phosphodiester group between the 2′-deoxyribose residues. The DNA tetrahedra containing TWJs were shown to undergo an unhooking reaction by the human DNA repair protein O⁶-alkylguanine DNA alkyltransferase (hAGT) resulting in structure disassembly. The unhooking reaction of hAGT towards the DNA tetrahedra was observed to be moderate to virtually complete depending on the protein equivalents. DNA tetrahedron structures have been explored as drug delivery platforms that release their payload in response to triggers, such as light, chemical agents or hybridization of release strands. The dismantling of DNA tetrahedron structures by a DNA repair protein contributes to the armamentarium of approaches for drug release employing DNA nanostructures.     

诺氟沙星与DNA的拉曼光谱研究

报道了诺氟沙星(NFX)及诺氟沙星胶囊内容物的FT-Raman光谱和在银胶基底上的表面增强拉曼光谱(SERS),归属了各个振动;研究了诺氟沙星与DNA的相互作用的SERS,结果表明:胶囊内容物的拉曼光谱图与对照品的拉曼光谱图的特征振动峰:C-F键的伸缩振动,C=C伸缩振动,O-C-O的对称伸缩振动峰值未发生变化,发生变化主要是分子骨架振动峰,诺氟沙星胶囊的辅料对拉曼光谱无实质影响,可建立拉曼光谱法检测诺氟沙星药物的分析方法;诺氟沙星可以在没有金属离子的存在下与DNA直接作用,与DNA相互作用的主要键合模式是插入作用,NFX分子中的平面结构插入DNA的双螺旋碱基平面,为深入了解喹诺酮类抗生素的抗菌机理提供可靠依据.     

Spectrum of the Micromaser with Kerr Medium

We have established the master equation for the micromaser with Kerr medium field density operator,studied the spectrum of the micromaser with Kerr medium and analyzed the influence of Kerr effect and the detuning on the spectrum.In the thermal-atom regime,we find that Kerr effect broadens Linewidth D and increases frequency-shift S,and that the detuning Δ narrows linewidth D and increases frequency-shift S as a whole,Moreover Kerr effect leads to oscillatings more rapidly in the resonance peaks,which means that it causes quantum noise,As a whole,with the increase of cavity-length L,the linewidth D and frequency-shift S gradually increase.