UV resonance Raman spectroscopy probes the localization of tryptophan‐containing antimicrobial peptides in lipid vesicles

In this work we employed UV resonance Raman spectroscopy with 229 nm excitation to study two tryptophan‐containing antimicrobial peptides with a broad‐spectrum activity against Gram‐positive and Gram‐negative bacteria: lactoferricin B (LfB, RRWQWRMKKLG) and pEM‐2 (KKWRWWLKALAKK). UV resonance Raman spectra of both peptides are dominated by tryptophan bands. Raman spectra of LfB and pEM‐2 in D₂O and 2,2,2‐trifluoro ethanol (TFE) have been measured and used to identify the hydrogen‐bond strength marker bands W6 and W17. The tryptophan doublet, W7, at 1340 and 1360 cm⁻¹ was used to detect an increase in the hydrophobicity of Trp environment in TFE. The spectra of LfB in complex with model cell membranes composed of zwitterionic dipalmitoylglycero‐phosphocholine (DPPC) or anionic dipalmitoyglycero‐phosphoglycerol (DPPG) lipid vesicles revealed a more hydrophobic Trp environment in DPPG, suggesting stronger interactions between the cationic peptide and anionic model cell membrane. Copyright © 2008 John Wiley & Sons, Ltd.     

Ultraviolet resonance Raman spectroscopy of a β‐sheet peptide: a model for membrane protein folding

The partitioning of a hydrophobic hexapeptide, N‐acetyl‐tryptophan‐pentaleucine (AcWL5), into self‐associated β‐sheets within a vesicle membrane was studied as a model for integral membrane protein folding and insertion via vibrational and electronic spectroscopy. Ultraviolet resonance Raman spectroscopy allows selective examination of the structures of amino acid side chains and the peptide backbone and provides information about local environment and molecular conformation. The secondary structure of AcWL5 within a vesicle membrane was investigated using 207.5‐nm excitation and found to consist of β‐sheets, in agreement with previous studies. The β‐sheet peptide shows enhanced Raman scattering cross‐sections for all amide modes as well as extensive hydrogen‐bonding networks. Tryptophan vibrational structure was probed using 230‐nm excitation. Increases in Raman cross‐sections of tryptophan modes W1, W3, W7, W10, W16, W17, and W18 of membrane‐incorporated AcWL5 are primarily attributed to greater resonance enhancement with the B_b electronic transition. The W17 mode, however, undergoes a much greater enhancement than is expected for a simple resonance effect, and this observation is discussed in terms of hydrogen bonding of the indole ring in a hydrophobic environment. The observed tryptophan mode frequencies and intensities overall support a hydrophobic environment for the indole ring within a vesicle, and these results have implications for the location of tryptophan in membrane protein systems. Copyright © 2009 John Wiley & Sons, Ltd.     

The effects of L‐ to D‐isomerization and C‐terminus deamidation on the secondary structure of antimicrobial peptide Anoplin in aqueous and membrane mimicking environment

UV resonance Raman spectra of the antimicrobial peptide (AMP) Anoplin (L ‐Anoplin‐NH₂) and two of its derivatives (enantiomer D ‐Anoplin‐NH₂ and C‐terminus deamidated L ‐Anoplin‐OH) were measured in aqueous buffer solution and in membrane‐mimetic environments including 2,2,2‐trifluoro ethanol (TFE), zwitterionic lipid dipalmitoylglycerophosphocholine (DPPC) and anionic lipid dipalmitoylglycerophosphoglycerol (DPPG) vesicle solutions. All three peptides were found to adopt random‐coil/β turn‐like conformation in aqueous solution over the temperature range of 1–60 °C. The conformation was found to become more α‐helical in membrane‐mimetic solutions such as TFE and DPPG but not in DPPC for all Anoplin derivatives. The data demonstrate that Anoplin preferentially binds to the anionic over the zwitterionic model cell membranes. Results also showed that deamidation does not change the conformation of L ‐Ano‐NH₂ very significantly, but does alter membrane rupturing and antimicrobial activities thus confirming that it is the physicochemical properties rather than the peptide conformation that define the mechanism of AMP action. Copyright © 2010 John Wiley & Sons, Ltd.     

Non‐invasive time‐course imaging of apoptotic cells by confocal Raman micro‐spectroscopy

Confocal Raman micro‐spectroscopy (CRMS) was used to measure time‐course spectral images of live cells undergoing apoptosis without using molecular labels or other invasive procedures. Human breast cancer cells (MDA‐MB‐231) were exposed to 300 µM etoposide to induce apoptosis, and Raman spectral images were acquired from the same cells at 2‐h intervals over a period of 6 h. The purpose‐built inverted confocal Raman micro‐spectrometer integrated an environmental enclosure and wide‐field fluorescence imaging. These key instrumental elements allowed the cells to be maintained under sterile physiological conditions (37 °C, 5% CO₂) and enabled viability and apoptosis assays to be carried out on the cells at the end of CRMS measurements. The time‐course spectral images corresponding to DNA Raman bands indicated an increase in signal intensity in apoptotic cells, which was attributed to DNA condensation. The Raman spectral images of lipids indicated a high accumulation of membrane phospholipids and highly unsaturated non‐membrane lipids in apoptotic cells. This study demonstrates the potential of CRMS for label‐free time‐course imaging of individual live cells. This technique may become a useful tool for in vitro toxicological studies and testing of new pharmaceuticals, as well as other time‐dependent cellular processes, such as cell differentiation, cell cycle and cell–cell interactions. Copyright © 2010 John Wiley & Sons, Ltd.     

Adsorption and reduction reactions of anthraquinone derivatives on gold electrodes studied with electrochemical surface‐enhanced Raman spectroscopy

Electrochemical surface‐enhanced Raman spectroscopy (EC‐SERS), combined with cyclic voltammetry, and the density functional theoretical (DFT) method were used to investigate self‐assembled monolayer (SAM) adsorption and reduction processes. Here, we choose the system of interest, being thiolacetyl‐terminated 2‐phenylene ethynylene‐substituted anthraquinone molecule (2‐AQ) on gold electrodes in buffered aqueous and aprotic solutions. In the buffered aqueous solution, the results of cyclic voltammetry and EC‐SERS measurements, as well as DFT calculations, indicate that the adsorbed molecules pass through a two‐electron two‐proton reduction reaction with cathodic polarization. In particular, the latter two methods confirmed the structural changes of SAMs during the process of redox reaction, 2‐AQ + 2e + 2H⁺ → 2‐AQH₂, where 2‐AQ and 2‐AQH₂ are the oxidized and reduced forms, respectively. In aprotic solutions (acetonitile), a stepwise reaction mechanism was proposed on the basis of the results of EC‐SERS and DFT calculations. The first reduction peak should be a half reaction process 2‐AQ + e → 2‐AQ⁻, where 2‐AQ⁻ is a single electron reduced form. Compared with that of 2‐AQ SAMs in the buffered aqueous solution, the results of EC‐SERS and DFT calculations in aprotic solution suggested that the solvent effect significantly influences the redox process of 2‐AQ in electrochemical interfaces. Copyright © 2012 John Wiley & Sons, Ltd.     

Vibrational dynamics of nitromethane mixed with IR780 dye studied by coherent anti‐stokes Raman spectroscopy

Vibrational coupling between different kinds of molecules in liquid mixture is studied by multiplex coherent anti‐Stokes Raman spectroscopy (CARS). To identify vibrational coherence, fs‐probe with high time resolution and narrowband‐probe with high spectral resolution are adopted in CARS experiments. Using liquid nitromethane (NM) mixed with organic dye IR780 perchlorate as the sample, we can clearly observe the interference between different vibrational modes. The intermolecular vibrational interaction between NM and IR780 molecules results in the vibrational coherence transfer (VCT) in the form of a change of phase correlation. Compared with symmetric bending vibration of NO₂, coherence transfer is found to be easier to take place between C―N bond of NM and vibrations of IR780, which indicates the selectivity of intermolecular vibrational interaction. The selectivity is deduced to be related to the coordination between intramolecular and collective motion of molecules. Copyright © 2016 John Wiley & Sons, Ltd.     

Quantitative detection of captopril in tablet and blood plasma samples by the combination of surface‐enhanced Raman spectroscopy with multiplicative effects model

With p‐thiocresol as internal standard, quantitative analysis of captopril, a synthetic angiotensin converting enzyme inhibitor, was achieved by the combination of the multiplicative effects model with surface‐enhanced Raman spectroscopy (SERS). The multiplicative effects model was adopted to correct the detrimental effects caused by the heterogeneity in the physical properties of enhancing substrate (i.e. Ag nano‐particles). Experimental results showed that the calibration model built on the SERS spectra of the calibration captopril samples prepared with ultrapure water could attain quite satisfactory concentration predictions for captopril in both real‐world tablet samples and plasma samples. The recovery rates were in the range of 94.3% to 109.8%, which were in substantial agreement with the corresponding results of LC‐MS/MS. The limit of detection and limit of quantification were estimated to be 0.149 and 0.451 μM, respectively. The proposed approach has advantages of relatively low cost, simplicity, high sensitivity and good accuracy and therefore can be further developed and extended to a routine method for the quantification of captopril in complex systems. Copyright © 2015 John Wiley & Sons, Ltd.     

膜系光谱系数对膜层参数的一阶和二阶偏导数的计算模型

为了寻找一种膜系深入分析与设计的有力辅助计算工具,从光学薄膜光谱系数的矩阵计算理论出发,基于对矩阵求迹运算的巧妙运用,从数学上建立了准确计算膜系光谱系数关于膜层几何厚度、实际折射率和消光系数等膜层参数的一阶和二阶偏导数的解析模型。这一偏导数计算模型物理上与矩阵理论具有一致的通用性和普适性,适用于任何各向同性的均匀膜系统。在数学上也严格成立,数值编程计算不存在差分近似,精度达到了计算机浮点运算的极限精度。而且算法运算费时少,计算速度快,取得了高度准确性和便于实时化的双重优越性能,非常有利于应用于膜系设计领域来提高膜层数较大时的设计速度和效率。同时,这一模型能非常便捷和准确地给出膜系许多实用的导数信息,对膜系分析、测量和膜厚监控等领域中的相关应用具有重要的参考意义。     

The structure of neutron-rich calcium isotopes studied by the shell model with realistic effective interactions

We study the structure of neutron-rich calcium isotopes in the shell model with realistic interactions. The CD-Bonn and Kuo-Brown (KB) interactions are used. As these interactions do not include the three-body force, their direct use leads to poor results. We tested whether the adjustment of the single particle energies (SPEs) would be sufficient to include the three-body correlations empirically. It turns out that the CD-Bonn interaction, after the adjustment of SPEs, gives good agreement with the experimental data for the energies and spectroscopy. For the KB interaction, both the SPEs and monopole terms require adjustments. Thus, the monopole problem is less serious for modern realistic interactions which include perturbations up to the third order. We also tested the effect of the non-central force on the shell structure. It is found that the effect of the tensor force in the CD-Bonn interaction is weaker than in the KB interaction.     

基于粒子群优化支持向量机的太阳电池温度预测

建立通用而精确的太阳电池热模型对光伏系统的建模、输出功率与转换效率的损失分析至关重要. 基于复杂的太阳电池温度机理, 分别研究了太阳电池温度的稳态热模型(steady state thermal model, SSTM)和支持向量机(support vector machines, SVM) 方法建立的精确预测热模型. 首先, 基于空气温度、太阳辐射强度、风速3个最主要因素与太阳电池温度的近似线性关系, 在已有SSTM的基础上, 建立并校正了太阳电池的SSTM并采用差分进化算法提取模型的未知参数. 其次, 为提高SVM的模型预测精度, 采用粒子群优化(particle swarm optimization, PSO) 算法对SVM的核参数和惩罚因子进行动态寻优, 在确定输入/输出样本集并划分训练集和测试集的基础上, 建立了基于粒子群优化支持向量机(PSO-SVM)的太阳电池温度精确预测热模型. 最后, 搭建实验平台, 在实验操作过程中减弱空气湿度、太阳入射角和热迟滞效应等因素对太阳电池温度的耦合. 通过实验对比表明, 建立的预测热模型性能可靠、全面、简洁, 其参数寻优算法优于遗传算法和交叉校验法, 模型预测精度优于反向传播神经网络(back propagation neural network) 和SSTM.