Effect of signal intensity normalization on the multivariate analysis of spectral data in complex ‘real‐world’ datasets

Spectral signal intensities, especially in ‘real‐world’ applications with nonstandardized sample presentation due to uncontrolled variables/factors, commonly require additional spectral processing to normalize signal intensity in an effective way. In this study, we have demonstrated the complexity of choosing a normalization routine in the presence of multiple spectrally distinct constituents by probing a dataset of Raman spectra. Variation in absolute signal intensity (90.1% of total variance) of the Raman spectra of these complex biological samples swamps the variation in useful signals (9.4% of total variance), degrading its diagnostic and evaluative potential. Using traditional spectral band choices, it is shown that normalization results are more complex than generally encountered in traditionally designed sample sets investigating limited chemical species. We demonstrate that no choice of a single band proves to be appropriate for predicting all the reference parameters, instead requiring a tailored normalization routine for each parameter. Of the reference parameters studied in the chosen system, signals from pathogenic adducts in ocular tissues called advanced glycation endproducts were most prominent when normalizing about the 1550–1690 cm⁻¹ region of the spectrum (17.5% of total variance, compared with 0.3% for unnormalized), while prediction of pentosidine and gender were optimized by normalization about the 1570 (R² = 0.97 vs 0.57 for unnormalized) and 1003 cm⁻¹ (p < 0.0000001 vs p < 0.01 for unnormalized) bands, respectively. The data obtained point to the extreme sensitivity of multivariate analysis to signal intensity normalization. Some general guidelines for making appropriate band choices are given, including the use of peak‐finding routines. 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.     

Thermal stability and photoluminescence property of hexagonal MoO3·0.55H2O microrods

The metastable phase of well-faceted, hexagonal, prism-like molybdenum oxide hydrate (MoO₃·0.55H₂O) was successfully synthesized by evaporating molybdic acid solution prepared through cation membrane electrolysis of Na₂MoO₄·2H₂O aqueous solution. The obtained crystals were characterized by X-ray diffraction (XRD), thermogravimetric (TG), scanning electronic microscopy (SEM) and photoluminescence (PL) spectrophotometry. The as-prepared MoO₃·0.55H₂O rods were of 2–4 μm in width and 5–12 μm in length. The MoO₃·0.55H₂O microrods displayed photoluminescence properties at room temperature and were transformed into stable orthorhombic α-MoO₃ after air annealing at 380 °C. Moreover, the influence of temperature factor on the phase transformation process, morphology and photoluminescence properties of MoO₃·0.55H₂O was investigated in detail.     

Application of high-resolution magic-angle spinning NMR spectroscopy to define the cell uptake of MRI contrast agents

A new method, based on proton high-resolution magic-angle spinning ((1)H HR-MAS) NMR spectroscopy, has been employed to study the cell uptake of magnetic resonance imaging contrast agents (MRI-CAs). The method was tested on human red blood cells (HRBC) and white blood cells (HWBC) by using three gadolinium complexes, widely used in diagnostics, Gd-BOPTA, Gd-DTPA, and Gd-DOTA, and the analogous complexes obtained by replacing Gd(III) with Dy(III), Nd(III), and Tb(III) (i.e., complexes isostructural to the ones of gadolinium but acting as shift agents). The method is based on the evaluation of the magnetic effects, line broadening, or induced lanthanide shift (LIS) caused by these complexes on NMR signals of intra- and extracellular water. Since magnetic effects are directly linked to permeability, this method is direct. In all the tests, these magnetic effects were detected for the extracellular water signal only, providing a direct proof that these complexes are not able to cross the cell membrane. Line broadening effects (i.e., the use of gadolinium complexes) only allow qualitative evaluations. On the contrary, LIS effects can be measured with high precision and they can be related to the concentration of the paramagnetic species in the cellular compartments. This is possible because the HR-MAS technique provides the complete elimination of bulk magnetic susceptibility (BMS) shift and the differentiation of extra- and intracellular water signals. Thus with this method, the rapid quantification of the MRI-CA amount inside and outside the cells is actually feasible.     

Role of non-lamellar-forming lipid in promotion of liposomal fusion

Membrane fusion is an important process in a wide range of cellular and sub-cellular activities. It is evident that during the intermediate stages of fusion some transitory non-bilayer configurations must appear within the lipid moiety. Using fluorescence techniques, we have studied here the process of aggregation and fusion of liposomes made of lipids, namely 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). When mixed together, the complete fusion between these two liposomes took around 44 h as both DPPC and DMPC favour lamellar configuration. When the mixture was incubated at 42°C the fusion process was completed after 23 h. But, when 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE) was added in the liposomal matrix the time for fusion was reduced to 21 h for mixture without incubation and 17 h when the mixture was incubated. This indicates that DPPE having a tendency to assume non-lamellar conformation, promoted destabilisation of the lamellar conformation within the liposome which facilitated the fusion between two apposing bilayers.     

PVDF微孔滤膜负载金纳米粒子用于牛奶中三聚氰胺的SERS快速检测

本文报道了一种利用聚偏二氟乙烯(Polyviny Lidene Fluoride,PVDF)微孔滤膜为载体的三聚氰胺(Melamine,MAM)的简便快速痕量分析技术。通过柠檬酸钠还原法制得平均粒径为30nm的纳米金溶胶,加入不同浓度的三聚氰胺后金纳米粒子快速聚集,这种纳米金聚集体可以通过简便快捷的过滤技术截留在PVDF微孔滤膜表面,进而用于SERS检测,在水溶液中最低检测浓度为0.05mg/L。我们进一步在牛奶样品中进行检测,在牛奶中检测限可以达到1mg/L,满足大部分国家规定的婴儿配方食品中三聚氰胺的限量值为1mg/kg的参考标准。该分析方法制备过程简单、成本低廉、总分析时间短,易于实现现场快速灵敏检测,应用前景广阔。     

Membrane lipid domains and dynamics as detected by Laurdan fluorescence

2-Dimethylamino-6-lauroylnaphthalene (Laurdan) is a membrane probe of recent characterization, which shows high sensitivity to the polarity of its environment. Steady-state Laurdan excitation and emission spectra have different maxima and shape in the two phospholipid phases, due to differences in the polarity and in the amount of dipolar relaxation. In bilayers composed of a mixture of gel and liquid-crystalline phases, the properties of Laurdan excitation and emission spectra are intermediate between those obtained in the pure phases. These spectral properties are analyzed using the generalized polarization (GP). TheGP value can be used for the quantitation of each phase. The wavelength dependence of theGP value is used to ascertain the coexistence of different phase domains in the bilayer. Moreover, by following the evolution of Laurdan emission vs. time after excitation, the kinetics of phase fluctuation in phospholipid vesicles composed of coexisting gel and liquid-crystalline phases was determined.GP measurements performed in several cell lines did not give indications of coexistence of phase domains in their membranes. In natural membranes, Laurdan parameters indicate a homogeneously fluid environment, with restricted molecular motion in comparison with the phospholipid liquid-crystalline phase. The influence of cholesterol on the phase properties of the two phospholipid phases is proposed to be the cause of the phase behavior observed in natural membranes. In bilayers composed of different phospholipids and various cholesterol concentrations, Laurdan response is very similar to that arising from cell membranes. In the absence of cholesterol, from the steady-state and time-resolved measurements of Laurdan in phospholipid vesicles, the condition for the occurrence of separate coexisting domains in the bilayer has been determined: the molecular ratio between the two phases must be in the range between 30% and 70%. Below and above this range, a single homogeneous phase is observed, with the properties of the more concentrated phase, slightly modified by the presence of the other. Moreover, in this concentration range, the calculated dimension of the domains is very small, between 20 and 50 Å.     

微观结构对膜换湿能力影响研究

对多孔膜换湿过程进行了理论分析,得到了透湿量和湿阻的计算式.同时对三种膜包括PVDF,PES和纤维素膜进行了湿阻的测量实验,通过对实验数据的回归处理,得到了多孔膜的结构参数.结果表明,膜厚、孔径分布、孔隙率及曲折因子对膜的换湿特性都有较大影响,在平均孔径相同的情况下,其它因素仍然导致了不同膜换湿能力的较大差距。在所获得的结构参数基础上,通过数值模拟的方法研究了平均孔径对膜换湿能力的影响。结果表明湿阻随平均孔径的增大而减小,减小趋势逐渐平缓最后趋向一定值。     

A novel design of membrane mirror with small deformation and imaging performance analysis in infrared system

A method of diminishing the shape error of membrane mirror is proposed in this paper. The inner inflating pressure is considerably decreased by adopting the pre-shaped membrane. Small deformation of the membrane mirror with greatly reduced shape error is sequentially achieved. Primarily a finite element model of the above pre-shaped membrane is built on the basis of its mechanical properties. Then accurate shape data under different pressures can be acquired by iteratively calculating the node displacements of the model. Shape data are applicable to build up deformed reflecting surfaces for the simulative analysis in ZEMAX. Finally, ground-based imaging experiments of 4-bar targets and nature scene are conducted. Experiment results indicate that the MTF of the infrared system can reach to 0.3 at a high spatial resolution of 10l p/mm, and texture details of the nature scene are well-presented. The method can provide theoretical basis and technical support for the applications in lightweight optical components with ultra-large apertures.     

Corona-induced graft polymerization for surface modification of porous polyethersulfone membranes

Graft polymerization of acrylic acid (AA) onto porous polyethersulfone (PES) membrane surfaces was developed using corona discharge in atmospheric ambience as an activation process followed by polymerization of AA in aqueous solution. The effects of the corona parameters and graft polymerization conditions on grafting yield (GY) of AA were investigated. The grafting of AA on the PES membranes was confirmed by ATR-FTIR and X-ray photoelectron spectroscopy (XPS) analysis. Porosimetry measurements indicate the average pore diameters and porosities of the modified membranes decrease with the increase of the GY. The hydrophilicity and surface wetting properties of the original and modified membranes were evaluated by observing the dynamic changes of water contact angles. It is found that the grafting of AA occurs not only on the membrane surfaces, but also on the pore walls of the cells inside the membrane. The permeability experiments of protein solution reveal that the grafting of PAA endows the modified membranes with enhanced fluxes and anti-fouling properties. The optimized GY of AA is in the range of 150-200 μg/cm². In addition, the tensile experiments show the corona discharge treatment with the power lower than 150 W yields little damage to the mechanical strength of the membranes.